US20130143757A1

US20130143757A1 - Methods and Compositions for Chlamydial Antigens as Reagents for Diagnosis of Tubal Factor Infertility and Chlamydial Infection

Info

Publication number: US20130143757A1
Application number: US13/691,260
Authority: US
Inventors: Guangming Zhong
Original assignee: University of Texas System
Current assignee: University of Texas System
Priority date: 2011-12-02
Filing date: 2012-11-30
Publication date: 2013-06-06
Also published as: US9046530B2

Abstract

The present invention provides Chlamydia proteins and fragments thereof and methods of use in diagnostic assays.

Description

STATEMENT OF PRIORITY

This application claims the benefit, under 35 U.S.C. §119(e), of U.S. Provisional Application Ser. No. 61/566,089, filed Dec. 2, 2011, the entire contents of which are incorporated by reference herein.

STATEMENT OF GOVERNMENT SUPPORT

The present invention was funded in part by government support under grant numbers R01AI064537 and R01AI047997 from the National Institutes of Health. The United States Government has certain rights in this invention.

STATEMENT REGARDING ELECTRONIC FILING OF A SEQUENCE LISTING

A Sequence Listing in ASCII text format, submitted under 37 C.F.R. §1.821, entitled 9237-50_ST25.txt, 1,098,928 bytes in size, generated on Nov. 29, 2012 and filed via EFS-Web, is provided in lieu of a paper copy. This Sequence Listing is hereby incorporated herein by reference into the specification for its disclosures.

BACKGROUND OF THE INVENTION

1. Field of the Invention
The present invention relates to the field of diagnosis/determination of chlamydial infection and disease as well as diagnosis/determination of tubal factor infertility.
2. Background Art
Twenty-five to 35% of patients presenting for infertility evaluation have tubal disease (1-4). Chlamydia trachomatis is the primary sexually transmitted infection responsible for tubal factor infertility (TFI) (5-7) with C. trachomatis antibodies in approximately 70% of human patients (8). C. trachomatis infected cells produce inflammatory cytokines (9-10) which may contribute to upper genital tract inflammatory damage (11-13). Lunefeld et al. found that among patients undergoing in vitro fertilization, those with C. trachomatis antibodies had decreased pregnancy rates (14).
C. trachomatis infection is often asymptomatic so patient history cannot dictate the presence of tubal disease (15-16). Elevated titers of anti-C. trachomatis antibodies are associated with TFI, but detection of overall antibody levels lacks the sensitivity and specificity required for differential diagnosis (17).
The present invention provides chlamydial antigens that can be used to develop rapid and convenient means for diagnosing/determining tubal factor infertility, as well as chlamydial antigens that can be used to develop rapid and convenient means for diagnosing/determining acute chlamydial invention.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a method of diagnosing tubal factor infertility in a subject, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof, and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and detecting the absence of formation of an antigen/antibody complex (e.g., detecting no antigen/antibody complex) with the antigen of Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof in the sample, thereby diagnosing tubal factor infertility in the subject.
Also provided herein is a method of identifying a subject as having an increased likelihood of having or developing tubal factor infertility, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and detecting the absence of formation of an antigen/antibody complex (e.g., detecting no antigen/antibody complex) with the antigen of Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof in the sample, thereby identifying the subject as having an increased likelihood of having or developing tubal factor infertility.
In further aspects, the present invention provides a method of diagnosing acute Chlamydia trachomatis infection in a subject, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby diagnosing acute Chlamydia trachomatis infection in the subject.
Additionally provided herein is a method of diagnosing acute Chlamydia trachomatis infection in a subject, comprising: (a) contacting a biological sample from the subject with an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof in the sample, thereby diagnosing acute Chlamydia trachomatis infection in the subject.
The present invention also provides a method of identifying a subject as having an increased likelihood of having an acute Chlamydia trachomatis infection, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby identifying the subject as having an increased likelihood of having an acute Chlamydia trachomatis infection.
Additionally provided herein is a method of identifying a subject as having an increased likelihood of having an acute Chlamydia trachomatis infection, comprising: (a) contacting a biological sample from the subject with an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof in the sample, thereby identifying the subject as having an increased likelihood of having an acute Chlamydia trachomatis infection.
Additionally provided herein is a method of identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection is likely to be or would be beneficial, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby identifying a subject for whom antibiotic therapy to treat Chlamydia trachomatis infection is likely to be or would be beneficial. The method above can further comprise the step of applying antibiotic therapy for Chlamydia trachomatis infection and/or anti-inflammatory treatment procedures to the subject.
Also provided herein is a method of identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection is likely to be or would be beneficial, comprising: (a) contacting a biological sample from the subject with an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof in the sample, thereby identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection is likely to be or would be beneficial. The method above can further comprise the step of applying antibiotic therapy for Chlamydia trachomatis infection and/or anti-inflammatory treatment procedures to the subject.
In further aspects, the present invention provides a kit comprising a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof.
Additionally provided is a kit comprising a diagnostic panel comprising an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof. In particular embodiments, a kit of this invention can further comprise an antigen of Chlamydia trachomatis CT147 or immunologically reactive fragment thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-F. Reactivity of 30 C. trachomatis Antigens with 54 Patient Sera at 1:4000 Dilution.

The 30 antigens were reacted with the 54 human sera as described in Table 2 except that each serum was diluted 1:4000 (data not shown). (A) Ten of the 30 antigens, failing to react with any of the 23 IFC sera, were thus presented in the figure. Note that HSP60 (CT110) and OmcB (CT443) maintained a detection sensitivity of 35.5% and 58%, respectively. (B) The reactivity of each of the 10 antigens was analyzed at individual antiserum level. Note that the combinations of CT443 with CT381 or HSP60 with CT376, CT381 and CT798 can have the highest sensitivity of 67.7% while maintaining 100% specificity. (C) The reactivity intensity between each antigen and the 21 positive sera (measured at individual antiserum level) was expressed as mean OD plus standard deviation, (D) Each of the 10 antigens was reacted with an antiserum sample pooled from the 21 sera at equal ratio without (D) or with absorption with C. trachomatis (CT)-infected HeLa lysate (E) or HeLa alone lysate (F). Note that absorption with CT-HeLa but not HeLa alone lysates removed the reactivity of each antigen with the pooled antiserum.

FIGS. 2A-B. Distribution Patterns of Reactivity of 5 Antigens with 24 TFI Samples.

The reactivity of each of 5 antigens (listed along the X-axis at the top of bottom) with 24 TFI and 25 FC human sera (each at a final dilution of 1:1000) was summarized in panel A and the reactivity with the 24 TFI samples (listed along Y-axis) was displayed in panel B. Each horizontal bar indicates a positive reactivity as determined based on the mean plus 2 standard deviations. Star indicates antisera uniquely reacted with a given antigen. Please note that CT110, CT376 & CT557 together positively detected a total of 14 unique TFI antisera (58%).

FIGS. 3A-C. Reactivity Patterns of CT443 and its 12 Fragments with TFI & FC Serum Samples.

CT443 full length (FL) and its 12 fragments (F1 to F12) as shown along the X-axis at the bottom were reacted with 24 TFI and 25 FC antisera (listed along the Y-axis on the left) at different dilutions (as shown along the Y-axis on the right), including 1:1,000 (A), 1:3,000 (B) and 1:10,000 (C). A positive reactivity was indicated with a horizontal bar. Number of or percentage of antisera reacted with a given antigen were spelled out in text in corresponding panels. Please note that most antibody reactivity was localized at the C-terminal fragments, including fragment 10 (F9), F11 and F12 and the fragments displayed better discrimination of TFI from FC samples than the full length CT443 at different serum dilutions.

FIGS. 4A-F. Reactivity Patterns of Antigens Preferentially Recognized by TFI Women with Serum Samples from 3 Different Groups of Women.

The reactivity of CT110, CT376, CT557, CT443 and CT443F11 as listed along the X-axis at the bottom with 24 TFI (A-B), 25 FC(C-D) and 24 STI (E-F) antisera as shown along the Y-axis at the left side. A positive reactivity was indicated with a horizontal bar. The human sera were used at either 1:1000 (A, C, E) or 1:10,000 (B, D, F) dilution. Antisera from the TFI group uniquely detected by a given antigen are marked with stars. Antigens preferentially recognized by TFI women were also highly reactive with STI women samples.

FIGS. 5A-B. Reactivity Patterns of 13 Antigens with STI Women Samples.

When the 21 antigens listed in Table 5 were reacted with human sera at 1:10,000 dilution, 13 antigens (listed at top and bottom of the figure) completely lost reactivity with either TFI or FC women samples but each maintained a certain level of reactivity with STI samples as summarized in panel A. The patterns of reactivity of the 13 antigens with 24 STI samples are shown in panel B. Each horizontal bar indicates a positive reactivity and antisera uniquely reacted with CT875 and CT147 are marked with a star. CT875 and CT147 detected 15 out of the 24 STI sera with a sensitivity of 63% while maintaining 100% specificity.

FIG. 6. Comparison of Chlamydia Antibody Test with Hysterosalpingogram (HSG) for Identifying Tubal Factor Infertility (TFI).

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based on the unexpected identification of immunodominant proteins of Chlamydia trachomatis and combinations thereof that allow for the diagnosis of tubal factor infertility as well as for the identification of subjects having an increased likelihood of having or developing tubal factor infertility. The present invention is also based on the unexpected identification of immunodominant proteins of Chlamydia trachomatis and combinations thereof that allow for the diagnosis of acute infection by Chlamydia trachomatis. These immunodominant proteins have been identified by the screening fusion protein arrays described herein in the EXAMPLES section. These immunodominant proteins, immunologically reactive fragments thereof and/or homologues of these proteins or immunologically reactive fragments thereof from other chlamydial species can be employed in methods of detection and diagnosis by identifying the presence of an antibody to the protein(s) and/or immunologically reactive fragment(s) thereof in a sample, such as a biological sample from a subject.
Thus, in one embodiment, the present invention provides a method of diagnosing tubal factor infertility in a subject, comprising, consisting essentially of or consisting of: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof, and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and not detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof in the sample, thereby diagnosing tubal factor infertility in the subject.
When a subject's infertility is diagnosed to be due to tubal factor as a result of chlamydial infection, a treatment option for the woman's infertility can be in vitro fertilization (IVF). Furthermore, applying both anti-chlamydial and anti-inflammatory treatment procedures to the subjects prior to IVF can increase the chance of success for IVF. Thus, in some embodiments, the method above can further comprise the step of advising and/or having the subject that has been diagnosed with tubal factor infertility to initiate IVF procedures to become pregnant. Such methods can further comprise the step of applying anti-chlamydial and/or anti-inflammatory treatment procedures to the subject.
Furthermore, the present invention provides a method of identifying a subject as having an increased likelihood of having or developing tubal factor infertility, comprising, consisting essentially of or consisting of: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and not detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof in the sample, thereby identifying the subject as having an increased likelihood of having or developing tubal factor infertility.
When a subject is identified as having an increased likelihood of having or developing tubal factor infertility as a result of chlamydial infection, the subject can undergo an infertility evaluation and both anti-chlamydial and anti-inflammatory treatment procedures can be applied to subject if the subject desires to have children. Thus, in some embodiments, the method above can further comprise the step of advising and/or having the subject that has been identified as having an increased likelihood of having or developing tubal factor infertility as a result of chlamydial infection to undergo an infertility evaluation. Such methods can further comprise the step of applying anti-chlamydial and/or anti-inflammatory treatment procedures to the subject.
Establishing that the cause of a woman's infertility is tubal factor infertility according to the methods described herein allows the woman and her physician to avoid approaches that are likely to be unsuccessful, such as hormone replacement and artificial insemination in order to overcome or bypass the woman's infertility and to select more effective methods of establishing a successful pregnancy in the woman, such as in vitro fertilization. The knowledge that the cause of a woman's infertility is tubal factor infertility also informs the woman and her physician that the use of invasive and costly procedures such as hysterosalpingogram (HSG) or laparoscopy may be unnecessary.
Thus, in one embodiment, the present invention also provides a method of identifying a subject as a good or suitable candidate for in vitro fertilization or surgical repair of tubal damage, comprising, consisting essentially of or consisting of: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and not detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof in the sample, thereby identifying the subject as a good or suitable candidate for in vitro fertilization or surgical repair of tubal damage. Thus, in some embodiments, the method above can further comprise the step of advising and/or having the subject that has been identified as a good or suitable candidate for in vitro fertilization (IVF) or surgical repair of tubal damage to undergo such IVF or surgery. Such methods can further comprise the step of applying anti-chlamydial and/or anti-inflammatory treatment procedures to the subject.
In some embodiments, the subject of this invention can be a human female and in some embodiments, the subject of this invention can be an infertile human female. In some embodiments, the tubal factor infertility can be caused by chlamydial infection and in some embodiments, the tubal factor infertility can be caused by infection by Chlamydia trachomatis.
In certain embodiments of the methods described above regarding tubal factor infertility, the diagnostic panel can further comprise, consist essentially of or consist of an additional antigen selected from the group consisting of a) Chlamydia trachomatis HSP60 or an immunologically reactive fragment thereof, b) Chlamydia trachomatis CT376 or an immunologically reactive fragment thereof, c) Chlamydia trachomatis CT557 or an immunologically reactive fragment thereof, and d) any combination thereof, and the method can further comprise, consist essentially of or consist of detecting an antigen/antibody complex in the sample for each of said additional antigen(s).
In further embodiments, the present invention provides a method of diagnosing acute Chlamydia trachomatis infection in a subject, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby diagnosing acute Chlamydia trachomatis infection in the subject. The method above can further comprise the step of applying anti-chlamydial and/or anti-inflammatory treatment procedures to the subject.
Also provided herein is a method of diagnosing acute Chlamydia trachomatis infection in a subject, comprising: (a) contacting a biological sample from the subject with an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof in the sample, thereby diagnosing acute Chlamydia trachomatis infection in the subject. The method above can further comprise the step of applying anti-chlamydial and/or anti-inflammatory treatment procedures to the subject.
The present invention also provides a method of identifying a subject as having an increased likelihood of having an acute Chlamydia trachomatis infection, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby identifying the subject as having an increased likelihood of having an acute Chlamydia trachomatis infection. The method above can further comprise the step of applying anti-chlamydial and/or anti-inflammatory treatment procedures to the subject.
Additionally provided herein is a method of identifying a subject as having an increased likelihood of having an acute Chlamydia trachomatis infection, comprising: (a) contacting a biological sample from the subject with an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof in the sample, thereby identifying the subject as having an increased likelihood of having an acute Chlamydia trachomatis infection. The method above can further comprise the step of applying anti-chlamydial and/or anti-inflammatory treatment procedures to the subject.
Additionally provided herein is a method of identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection is likely to be or would be beneficial, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby identifying a subject for whom antibiotic therapy to treat Chlamydia trachomatis infection is likely to be or would be beneficial. The method above can further comprise the step of applying antibiotic therapy for Chlamydia trachomatis infection and/or anti-inflammatory treatment procedures to the subject.
Also provided herein is a method of identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection is likely to be or would be beneficial, comprising: (a) contacting a biological sample from the subject with an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof in the sample, thereby identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection is likely to be or would be beneficial. The method above can further comprise the step of applying antibiotic therapy for Chlamydia trachomatis infection and/or anti-inflammatory treatment procedures to the subject.
In some embodiments the method of diagnosing acute chlamydial infection or identifying a subject as having an increased likelihood of having an acute Chlamydia trachomatis infection or identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection can further comprise, consist essentially of or consist of contacting the sample with an antigen of Chlamydia trachomatis CT147 or immunologically reactive fragment thereof and detecting an antigen/antibody complex in the sample with the antigen of Chlamydia trachomatis CT147 or immunologically reactive fragment thereof.
A biological sample of this invention can be any biological fluid and/or tissue in which antibodies can be detected. Nonlimiting examples of a sample of this invention can include vaginal fluid, vaginal tissue, vaginal washing, vaginal swab, vaginal discharge, cervical swab, cervical tissue urethral swab, urethral discharge, rectal swab, rectal material, rectal washing, urine, blood, serum, plasma, saliva, tears, skin swab, semen, seminal fluid, sputum, bronchial fluid, bronchial washing, peritoneal fluid, peritoneal washing, pleural fluid, pleural washing, cerebrospinal fluid, eye fluid and/or tissue, fluid and/or tissue from lung, liver, heart, brain, kidney, spleen or muscle and any combination thereof.
In some embodiments, the biological sample of this invention to be used in the methods of this invention can be diluted 1:10, 1:100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, 1:1000, 1:1500, 1:2000, 1:3000, 1:4000, 1:5000, 1:6000, 1:7000, 1:8000, 1:9000, 1:10,000, 1:20,000, 1:30,000, 1:40,000, 1:50,000, 1:100,000, etc. Such a dilution can be carried out according to protocols well known in the art and as described in the EXAMPLES section herein. Such dilution can be used to increase the specificity of the method, as described herein.
In the methods of this invention, the biological sample can be preabsorbed, e.g., to reduce or minimize cross-reactivity and/or background. As nonlimiting examples, in some embodiments, the biological sample can be preabsorbed with a lysate of bacteria expressing glutathione-S-transferase (GST) and/or a lysate of normal (e.g., non-chlamydial infected mammalian cells. In some embodiments absorption of the sample can be with a lysate of Chlamydia-infected mammalian cells, to remove and/or block chlamydial antigen-specific antibodies from human samples, which can help confirm the specificity of human antibody binding to the test antigen.
A subject of this invention that has an “increased likelihood” or “increased risk” of having or tubal factor infertility can be a subject having symptoms and/or signs of infertility and/or meets criteria known in the art for being infertile or such a subject can be a subject who is not having symptoms and/or signs of infertility and/or does not meet criteria know in the art for being infertile. By “increased likelihood” or “increased risk” of having tubal factor infertility it is meant that the increase is relative to a control (e.g., a subject whose biological sample is not positive for antibodies to CT443 and CT381 and negative for antibodies to CT875).
Furthermore, a subject of this invention that has an “increased likelihood” or “increased risk” of having an acute Chlamydia trachomatis infection can be a subject having symptoms and/or signs of Chlamydia trachomatis infection or such a subject can be a subject who is not having symptoms and/or signs of Chlamydia trachomatis infection. By “increased likelihood” or “increased risk” of acute Chlamydia trachomatis infection it is meant that the increase is relative to a control (e.g., a subject whose biological sample is not positive for antibodies to CT443 and CT381 and CT875 or a subject whose biological sample is not positive for antibodies to CT875).
In additional embodiments of this invention, the chlamydial proteins listed herein can also be employed in the methods and compositions of this invention, either singly or in any combination with one another and/or in combination with any other chlamydial protein and/or reagent of this invention.
As used herein, “a,” “an” or “the” can mean one or more than one. For example, “a” cell can mean a single cell or a multiplicity of cells.
Also as used herein, “and/or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”).
Furthermore, the term “about,” as used herein when referring to a measurable value such as an amount of a compound or agent of this invention, dose, time, temperature, and the like, is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified amount.
The term “isolated” as used herein means the protein or polypeptide or immunologically reactive fragment or nucleic acid of this invention is sufficiently free of contaminants or cell components with which polypeptides and/or nucleic acids normally occur. “Isolated” does not mean that the preparation is technically pure (homogeneous), but it is sufficiently pure to provide the polypeptide or nucleic acid in a form in which it can be used in methods of this invention.
The term “epitope” as used herein refers to at least about 3 to about 5, or about 5 to about 10 or about 5 to about 15, and not more than about 1,000 amino acids (or any integer therebetween) (e.g., 5-12 amino acids or 3-10 amino acids or 4-8 amino acids or 6-15 amino acids, etc.), which define a sequence that by itself or as part of a larger sequence, binds to an antibody generated in response to such sequence or stimulates a cellular immune response. There is no critical upper limit to the length of the fragment, which can comprise nearly the full-length of the protein sequence, or even a fusion protein comprising two or more epitopes from a single or multiple chlamydial proteins. An epitope for use in the subject invention is not limited to a polypeptide having the exact sequence of the portion of the parent protein from which it is derived. Indeed, there are many known strains or isolates of Chlamydia and there are several variable domains that exhibit relatively high degrees of variability between isolates. Thus, the term “epitope” encompasses sequences identical to the native sequence, as well as modifications to the native sequence, such as deletions, additions and substitutions (generally, but not always, conservative in nature). In some embodiments, an “immunologically reactive fragment” of this invention can comprise one, two, three, four or more epitopes of a protein of this invention.
Regions of a given polypeptide or fragment thereof that include an epitope can be identified using any number of epitope mapping techniques, well known in the art. (See, e.g., Epitope Mapping Protocols in Methods in Molecular Biology, Vol. 66, Glenn E. Morris, Ed., 1996, Humana Press, Totowa, N.J.). For example, linear epitopes can be determined by e.g., concurrently synthesizing large numbers of peptides on solid supports, the peptides corresponding to portions of the protein molecule, and reacting the peptides with antibodies while the peptides are still attached to the supports. Such techniques are known in the art and described in, e.g., U.S. Pat. No. 4,708,871; Geysen et al. (1984) Proc. Natl. Acad. Sci. USA 81:3998-4002; Geysen et al. (1986) Molec. Immunol. 23:709-715, all incorporated herein by reference in their entireties.
Similarly, conformational epitopes are readily identified by determining spatial conformation of amino acids such as by, e.g., x-ray crystallography and 2-dimensional nuclear magnetic resonance. Antigenic regions of proteins can also be identified using standard antigenicity and hydropathy plots, such as those calculated using, e.g., the Omiga version 1.0 software program available from the Oxford Molecular Group. This computer program employs the Hopp/Woods method (Hopp et al., Proc. Natl. Acad. Sci USA (1981) 78:3824-3828) for determining antigenicity profiles and the Kyte-Doolittle technique (Kyte et al., J. Mol. Biol. (1982) 157:105-132) for hydropathy plots.
As used herein, the term “polypeptide” or “protein” is used to describe a chain of amino acids that correspond to those encoded by a nucleic acid. A polypeptide or protein of this invention can be a peptide, which usually describes a chain of amino acids of from two to about 30 to about 50 amino acids. The term polypeptide as used herein also describes a chain of amino acids having more than about 30 amino acids or more than about 50 amino acids and can be a fragment or domain of a protein or a full length protein. Furthermore, as used herein, the term polypeptide can refer to a linear chain of amino acids or it can refer to a chain of amino acids that has been processed and folded into a functional protein. It is understood, however, that 30 or 50 is an arbitrary number with regard to distinguishing peptides and polypeptides and the terms can be used interchangeably for a chain of amino acids. The polypeptides of the present invention are obtained by isolation and purification of the polypeptides from cells where they are produced naturally, by enzymatic (e.g., proteolytic) cleavage, and/or recombinantly by expression of nucleic acid encoding the polypeptides or fragments of this invention. The polypeptides and/or fragments of this invention can also be obtained by chemical synthesis or other known protocols for producing polypeptides and fragments.
The amino acid sequences of this invention are presented in the amino to carboxy direction, from left to right. The “carboxy terminus” or “C terminus” of a protein or amino acid sequence as used herein refers to a portion or fragment or domain of a protein or amino acid sequence that makes up about ⅔, about ½, about ⅓ or about ¼ of the total amino acid sequence at the carboxy end of the sequence (i.e., the right end or right-sided end or the “end” of the sequence). The “amino terminus” or “N terminus” of a protein or amino acid sequence as used herein refers to a portion or fragment or domain of a protein or amino acid sequence that makes up about ⅔, about ½, about ⅓ or about ¼ of the total amino acid sequence at the amino end of the sequence (i.e., the left end or left sided end or the “beginning” of the sequence).
Nucleotide sequences are presented herein by single strand only, in the 5′ to 3′ direction, from left to right. However, it is intended that the nucleic acids of this invention can be either single or double stranded (i.e., including the complementary nucleic acid). A nucleic acid of this invention can be the complement of a nucleic acid described herein.
A “biologically active fragment” includes a polypeptide or peptide of this invention that comprises a sufficient number of amino acids to have one or more of the biological activities of the polypeptides of this invention. Such biological activities can include, but are not limited to, in any combination, binding activity and/or immunogenic activity, as well as any other activity now known or later identified for the polypeptides and/or fragments of this invention.
An “immunologically reactive fragment,” “immunogenic fragment” or “antigenic fragment” of a protein refers to a portion of the protein or peptide that is immunologically reactive with a binding partner, e.g., an antibody, which is immunologically reactive with the protein or peptide itself.
In some embodiments, the terms “immunologically reactive fragment,” “immunogenic fragment” or “antigenic fragment” are used to describe a fragment or portion of a protein or peptide that can stimulate a humoral and/or cellular immune response in a subject. An immunologically reactive fragment, immunogenic fragment or antigenic fragment of this invention can comprise, consist essentially of and/or consist of one, two, three, four or more epitopes of a protein of this invention. An immunologically reactive fragment, immunogenic fragment or antigenic fragment can be any fragment of contiguous amino acids of a Chlamydia trachomatis protein of this invention, including but not limited to CT443, CT381, CT875, CT147, HSP60, CT376, CT557, CT858 (CPAF), Pgp3, CT823 (cHtrA), CT681 (MOMP), CT119 (IncA), CT813, CT795, CT621 and CT622, the amino acid sequences of each of which are provided herein and are available at www.ncbi.nlm.nih.gov/protein/15605169) and can be for example, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 amino acids in length, dependent upon the total number of amino acids of the full length protein. Identification of any such immunologically reactive, immunogenic or antigenic fragment(s) is routine in the art.
Nonlimiting examples of an immunologically reactive fragment of Chlamydia trachomatis CT443 protein (GenBank Database® Accession No. NP_—219955.1) include amino acids 41-269 (F1), amino acids 166-269 (F2), amino acids 211-269 (F3), amino acids 101-210 (F4), amino acids 166-210 (F5), amino acids 41-165 (F6), amino acids 101-165 (F7), amino acids 41-100 (F8), amino acids 211-410 (F9), amino acids 270-410 (F10), amino acids 270-553 (F11) and amino acids 411-553 (F12), with amino acid numbering starting with amino acid 1 and ending with amino acid 553 of the 553 amino acid CT443 protein, the amino acid sequence of which is provided herein. As described herein, in some embodiments, fragments identified above as F1 through F8 could be described as N terminal or amino terminal fragments. As also described herein, in some embodiments, fragments identified above as F9 through F12 could be described as C terminal or carboxy terminal fragments. In some embodiments of this invention, the CT443 protein or immunologically reactive fragment employed in methods of this invention can be a C terminal fragment.
Nonlimiting examples of an immunologically reactive fragment of Chlamydia trachomatis CPAF protein (GenBank Database® Accession No. AAC68456.1) include amino acids 1-200, amino acids 136-609, amino acids 242-609, amino acids 284-609 and amino acids 387-609 with numbering starting from amino acid 1 at the amino terminus through amino acid 609 at the carboxy terminus of the 609 amino acid CPAF protein, the amino acid sequence of which is provided herein and is available under GenBank Database® Accession No. AAC68456.1.
Also provided herein is an isolated peptide comprising, consisting essentially of or consisting of about five amino acids to about 15, 20, 25, 30, 35, 40, 45, 50, 50, 70, 80, 90 or 100 amino acids (including any value between 5 and 100 not explicitly recited herein), wherein the peptide comprises one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) of the 5 mer peptides listed in Table 6, as well as a composition comprising any of these isolated peptides, singly or in any combination in a carrier (e.g., a pharmaceutically acceptable carrier). Such peptides can be employed as immunologically reactive fragments in the methods of this invention.
It is further provided herein that in the methods of this invention, an immunologically reactive fragment of this invention and a biological sample diluted as described herein can be employed in any combination in the methods described herein to increase the specificity of the method. As one nonlimiting example, a biological sample diluted 1:3000 was reacted with the F11 fragment of chlamydial protein CT443 resulting in an increase in specificity of the assay and further diluting the sample 1:10,000 increased specificity even more (see Example 2). Optimization of the sample dilution and immunologically reactive fragment reactivity can be carried out as described herein and according to methods well known in the art.
A fragment of a polypeptide or protein of this invention can be produced by methods well known and routine in the art. Fragments of this invention can be produced, for example, by enzymatic or other cleavage of naturally occurring peptides or polypeptides or by synthetic protocols that are well known. Such fragments can be tested for one or more of the biological activities of this invention according to the methods described herein, which are routine methods for testing activities of polypeptides, and/or according to any art-known and routine methods for identifying such activities. Such production and testing to identify biologically active fragments and/or immunologically reactive fragments of the polypeptides described herein would be well within the scope of one of ordinary skill in the art and would be routine.
As used herein, the term “antibody” includes intact immunoglobin molecules as well as fragments thereof, such as Fab, F(ab′)2, and Fc, which are capable of binding the epitopic determinant of an antigen (i.e., antigenic determinant). Antibodies that bind the polypeptides of this invention are prepared using intact polypeptides or fragments containing small peptides of interest as the immunizing antigen. The polypeptide or fragment used to immunize an animal can be derived from enzymatic cleavage, recombinant expression, isolation from biological materials, synthesis, etc., and can be conjugated to a carrier protein, if desired. Commonly used carriers that are chemically coupled to peptides and proteins for the production of antibody include, but are not limited to, bovine serum albumin, thyroglobulin and keyhole limpet hemocyanin. The coupled peptide or protein is then used to immunize the animal (e.g., a mouse, rat, or rabbit). The polypeptide or peptide antigens can also be administered with an adjuvant, as described herein and as otherwise known in the art. The term “antibody” as used herein, includes, but is not limited to a polypeptide encoded by an immunoglobulin gene or immunoglobulin genes, or a fragment thereof. “Antibody” also includes, but is not limited to, a polypeptide encoded by an immunoglobulin gene or immunoglobulin genes, or a fragment thereof, which specifically binds to and recognizes the biomarkers of this invention.
The term “epitope” means an antigenic determinant that is specifically bound by an antibody. Epitopes usually consist of surface groupings of molecules such as amino acids and/or sugar side chains and usually have specific three-dimensional structural characteristics, as well as specific charge characteristics.
The terms “specifically binds to” and “specifically reactive with” refer to a binding reaction that is determinative of the presence of the antigen and antibody or aptamer and target in the presence of a heterogeneous population of proteins, nucleic acids and/or other biologics. Thus, under designated assay conditions, the specified antibodies and antigens and/or aptamers and targets bind to one another and do not bind in a significant amount to other components present in a sample.
In some embodiments employing antibodies, a variety of immunoassay formats can be used to select antibodies specifically reactive with a particular antigen. For example, solid-phase ELISA immunoassays are routinely used to select monoclonal antibodies specifically immunoreactive with an analyte. See Harlow and Lane (ANTIBODIES: A LABORATORY MANUAL, Cold Springs Harbor Publications, New York, (1988)) for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity. Typically a specific or selective reaction will be at least twice background signal to noise and more typically more than 10 to 100 times greater than background.
Various immunoassays can be used to detect antibodies of this invention. Such immunoassays typically involve the measurement of antigen/antibody complex formation between a protein or peptide and its specific antibody.
The immunoassays of the invention can be either competitive or noncompetitive and both types of assays are well-known and well-developed in the art. In competitive binding assays, antigen or antibody competes with a detectably labeled antigen or antibody for specific binding to a capture site bound to a solid surface. The concentration of labeled antigen or antibody bound to the capture agent is inversely proportional to the amount of free antigen or antibody present in the sample.
Noncompetitive assays of this invention can be sandwich assays, in which, for example, the antigen is bound between two antibodies. One of the antibodies is used as a capture agent and is bound to a solid surface. The other antibody is labeled and is used to measure or detect the resultant antigen/antibody complex by e.g., visual or instrument means. A number of combinations of antibody and labeled antibody can be used, as are well known in the art. In some embodiments, the antigen/antibody complex can be detected by other proteins capable of specifically binding human immunoglobulin constant regions, such as protein A, protein L or protein G. These proteins are normal constituents of the cell walls of streptococcal bacteria. They exhibit a strong nonimmunogenic reactivity with immunoglobulin constant regions from a variety of species. (See, e.g., Kronval et al., J. Immunol., 111:1401-1406 (1973); Akerstrom et al., J. Immunol., 135:2589-2542 (1985).)
In some embodiments, the non-competitive assays need not be sandwich assays. For instance, the antibodies or antigens in the sample can be bound directly to the solid surface. The presence of antibodies or antigens in the sample can then be detected using labeled antigen or antibody, respectively.
In some embodiments, antibodies and/or proteins can be conjugated or otherwise linked or connected (e.g., covalently or noncovalently) to a solid support (e.g., bead, plate, slide, dish, membrane or well) in accordance with known techniques. Antibodies can also be conjugated or otherwise linked or connected to detectable groups such as radiolabels (e.g., ³⁵S, ¹²⁵I, ³²P, ¹³H, ¹⁴C, ¹³¹I), enzyme labels (e.g., horseradish peroxidase, alkaline phosphatase), gold beads, chemiluminescence labels, ligands (e.g., biotin) and/or fluorescence labels (e.g., fluorescein) in accordance with known techniques.
A variety of organic and inorganic polymers, both natural and synthetic can be used as the material for the solid surface. Nonlimiting examples of polymers include polyethylene, polypropylene, poly(4-methylbutene), polystyrene, polymethacrylate, poly(ethylene terephthalate), rayon, nylon, poly(vinyl butyrate), polyvinylidene difluoride (PVDF), silicones, polyformaldehyde, cellulose, cellulose acetate, nitrocellulose, and the like. Other materials that can be used include, but are not limited to, include paper, glass, ceramic, metal, metalloids, semiconductive materials, cements and the like. In addition, substances that form gels, such as proteins (e.g., gelatins), lipopolysaccharides, silicates, agarose and polyacrylamides can be used. Polymers that form several aqueous phases, such as dextrans, polyalkylene glycols or surfactants, such as phospholipids, long chain (12-24 carbon atoms) alkyl ammonium salts and the like are also suitable. Where the solid surface is porous, various pore sizes can be employed depending upon the nature of the system.
A variety of immunoassay systems can be used, including but not limited to, radio-immunoassays (RIA), enzyme-linked immunosorbent assays (ELISA) assays, enzyme immunoassays (EIA), “sandwich” assays, gel diffusion precipitation reactions, immunodiffusion assays, agglutination assays, immunofluorescence assays, fluorescence activated cell sorting (FACS) assays, immunohistochemical assays, protein A immunoassays, protein G immunoassays, protein L immunoassays, biotin/avidin assays, biotin/streptavidin assays, immunoelectrophoresis assays, precipitation/flocculation reactions, immunoblots (Western blot; dot/slot blot); immunodiffusion assays; liposome immunoassay, chemiluminescence assays, library screens, expression arrays, etc., immunoprecipitation, competitive binding assays and immunohistochemical staining. These and other assays are described, among other places, in Hampton et al. (Serological Methods, a Laboratory Manual, APS Press, St Paul, Minn. (1990)) and Maddox et al. (J. Exp. Med. 158:1211-1216 (1993); the entire contents of which are incorporated herein by reference for teachings directed to immunoassays).
The methods of this invention can also be carried out using a variety of solid phase systems, such as described in U.S. Pat. No. 5,879,881, as well as in a dry strip lateral flow system (e.g., a “dipstick” system), such as described, for example, in U.S. Patent Publication No. 20030073147, the entire contents of each of which are incorporated by reference herein.
The term “antibody” or “antibodies” as used herein refers to all types of immunoglobulins, including IgG, IgM, IgA, IgD, and IgE. The antibody can be monoclonal or polyclonal and can be of any species of origin, including, for example, mouse, rat, rabbit, horse, goat, sheep or human, or can be a chimeric or humanized antibody. See, e.g., Walker et al., Molec. Immunol. 26:403-11 (1989). The antibodies can be recombinant monoclonal antibodies produced according to the methods disclosed in U.S. Pat. No. 4,474,893 or U.S. Pat. No. 4,816,567. The antibodies can also be chemically constructed according to the method disclosed in U.S. Pat. No. 4,676,980. The antibody can further be a single chain antibody or bispecific antibody.
Antibody fragments included within the scope of the present invention include, for example, Fab, F(ab′)2, and Fc fragments, and the corresponding fragments obtained from antibodies other than IgG. Such fragments can be produced by known techniques. For example, F(ab′)2 fragments can be produced by pepsin digestion of the antibody molecule, and Fab fragments can be generated by reducing the disulfide bridges of the F(ab′)2 fragments. Alternatively, Fab expression libraries can be constructed to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity (Huse et al., (1989) Science 254:1275-1281).
Monoclonal antibodies can be produced in a hybridoma cell line according to the technique of Kohler and Milstein, (1975) Nature 265:495-97. For example, a solution containing the appropriate antigen can be injected into a mouse and, after a sufficient time, the mouse sacrificed and spleen cells obtained. The spleen cells are then immortalized by fusing them with myeloma cells or with lymphoma cells, typically in the presence of polyethylene glycol, to produce hybridoma cells. The hybridoma cells are then grown in a suitable medium and the supernatant screened for monoclonal antibodies having the desired specificity. Monoclonal Fab fragments can be produced in bacterial cell such as E. coli by recombinant techniques known to those skilled in the art. See, e.g., W. Huse, (1989) Science 246:1275-81.
Antibodies can also be obtained by phage display techniques known in the art or by immunizing a heterologous host with a cell containing an epitope of interest.
The term “sample” as used herein is used in its broadest sense. A biological sample suspected of containing a polypeptide, fragment, antibody and/or nucleic acid of this invention can be any biological fluid, an extract from a cell, an extracellular matrix isolated from a cell, a cell (in solution or bound to a solid support), a tissue, a tissue print, and the like.
A “pharmaceutically acceptable” component such as a salt, carrier, excipient or diluent of a composition according to the present invention is a component that (i) is compatible with the other ingredients of the composition in that it can be combined with the compositions of the present invention without rendering the composition unsuitable for its intended purpose, and (ii) is suitable for use with subjects as provided herein without undue adverse side effects (such as toxicity, irritation, and allergic response). Side effects are “undue” when their risk outweighs the benefit provided by the composition. Non-limiting examples of pharmaceutically acceptable components include, without limitation, any of the standard pharmaceutical carriers such as phosphate buffered saline solutions, water, emulsions such as oil/water emulsion, microemulsions and various types of wetting agents.
A variety of protocols for detecting the presence of and/or measuring the amount of an antibody or antibodies in a sample are known in the art. Such protocols are well known in the art and non-limiting examples include enzyme immunoassays (ETA), agglutination assays, immunoblots (Western blot; dot/slot blot, etc.), radioimmunoassays (RIA), immunodiffusion assays, chemiluminescence assays, antibody library screens, expression arrays, enzyme-linked immunosorbent assays (ELISA), radioimmunoassays (RIA), immunoprecipitation, Western blotting, competitive binding assays, immunofluorescence, immunohistochemical staining precipitation/flocculation assays and fluorescence-activated cell sorting (FACS). These and other assays are described, among other places, in Hampton et al. (Serological Methods, a Laboratory Manual, APS Press, St Paul, Minn. (1990)) and Maddox et al. (J. Exp. Med. 158:1211-1216 (1993)).
The present invention further provides isolated polypeptides, peptides, proteins and/or fragments that are substantially equivalent to those described for this invention. As used herein, “substantially equivalent” can refer both to nucleic acid and amino acid sequences, for example a mutant sequence, that varies from a reference sequence by one or more substitutions (e.g., substitution with conservative amino acids as are well known in the art), deletions and/or additions, the net effect of which does not result in an undesirable adverse functional dissimilarity between reference and subject sequences. In some embodiments, this invention can include substantially equivalent sequences that have an adverse functional dissimilarity. For purposes of the present invention, sequences having equivalent biological activity and equivalent expression characteristics are considered substantially equivalent.
The invention further provides homologues, as well as methods of obtaining homologues, of the polypeptides and/or fragments of this invention from other strains of Chlamydia and/or other organisms included in this invention. As used herein, an amino acid sequence or protein is defined as a homologue of a polypeptide or fragment of the present invention if it shares significant homology to one of the polypeptides and/or fragments of the present invention. Significant homology means at least 75%, 80%, 85%, 90%, 95%, 98% and/or 100% homology with another amino acid sequence. Specifically, by using the nucleic acids that encode the chlamydial proteins of this invention (as are known in the art and incorporated by reference herein), as a probe or primer, and techniques such as PCR amplification and colony/plaque hybridization, one skilled in the art can identify homologues of the polypeptides and/or fragments of this invention in Chlamydia and/or other organisms on the basis of information available in the art. A homologue of a Chlamydia trachomatis protein of this invention can include a protein of Chlamydia muridarum, Chlamydia pneumoniae, Chlamydia psittaci or Chlamydia caviae identified to be a homologue according to methods well known in the art and as described herein. It would be well understood by one of ordinary skill in the art that such homologues (either complete proteins and/or immunologically reactive fragments thereof) can be employed in the methods of this invention. The identification and testing of such homologues for suitability as antigens in the methods of this invention is well within the skill of one in the art. Such homologues among the proteins of Chlamydia trachomatis, Chlamydia muridarum, Chlamydia pneumoniae, Chlamydia psittaci and Chlamydia caviae are well known in the art. As one non-limiting example, a listing of Chlamydia pneumoniae proteins and the Chlamydia trachomatis homologues of these proteins can be found in U.S. Pat. No. 6,822,071, the entire contents of which are incorporated by reference herein for these teachings.
In some embodiments, the present invention provides the antigens of this invention immobilized on a solid support (e.g., beads, plates, slides or wells formed from materials such as, e.g., latex or polystyrene). Nonlimiting examples of such solid supports include polycarbonate, agarose, nitrocellulose, sepharose, acrylic resins, polyacrylamide and latex beads, as well as any other solid support known in the art. Techniques for coupling antibodies and antigens to such solid supports are well known in the art (Weir et al., Handbook of Experimental Immunology 4th Ed., Blackwell Scientific Publications, Oxford, England, Chapter 10 (1986)). Antibodies and/or antigens of this invention can likewise be conjugated to detectable groups such as radiolabels (e.g., ³⁵S, ¹²⁵I, ¹³¹I), enzyme labels (e.g., horseradish peroxidase, alkaline phosphatase), and fluorescence labels (e.g., fluorescein) in accordance with known techniques. Conditions suitable for the formation of an antigen/antibody complex are routine in the art and form the basis for all immunoassays. Such conditions may vary depending on the particular reagents, samples and/or steps employed in a given immunoassay, as would be readily determined by one of ordinary skill in the art. Determination of the formation of an antibody/antigen complex in the methods of this invention can be by detection of, for example, precipitation, agglutination, flocculation, radioactivity, color development or change, fluorescence, luminescence, etc., as is well know in the art.
It is further contemplated that the present invention provides kits for detection, in a biological sample, of antibodies specifically reactive to antigens of this invention. In one embodiment, the kit can comprise one or more antigens of this invention, along with suitable buffers, wash solutions and/or other reagents for the detection of antibody/antigen complex formation.
In one embodiment, a kit of this invention can comprise, consist essentially of and/or consist of antigens (e.g., a diagnostic panel) and reagents for detecting, in a biological sample, the presence or absence of 1) an antibody that specifically reacts with Chlamydia trachomatis CT443 protein or an immunologically reactive fragment thereof, 2) an antibody that specifically reacts with Chlamydia trachomatis CT875 protein or an immunologically reactive fragment thereof and 3) Chlamydia trachomatis CT381 protein or an immunologically reactive fragment thereof. In some embodiments, the kit described in this paragraph can further comprise, consist essentially of or consist of antigens (e.g., as additional components of a diagnostic panel) and reagents for detecting, in a biological sample, the presence or absence of Chlamydia trachomatis HSP60 protein or an immunologically reactive fragment thereof, Chlamydia trachomatis CT376 protein or an immunologically reactive fragment thereof, Chlamydia trachomatis CT557 protein or an immunologically reactive fragment thereof, and any combination thereof. Thus, a kit of this invention can include a diagnostic panel that comprises, consists essentially of or consists of an antigen of Chlamydia trachomatis CT443 protein or an immunologically reactive fragment thereof, an antigen of Chlamydia trachomatis CT875 protein or an immunologically reactive fragment thereof, an antigen of Chlamydia trachomatis CT381 protein or an immunologically reactive fragment thereof. Such a kit can further include, in its diagnostic panel, an antigen of Chlamydia trachomatis HSP60 protein or an immunologically reactive fragment thereof, an antigen of Chlamydia trachomatis CT376 protein or an immunologically reactive fragment thereof, an antigen of Chlamydia trachomatis CT557 protein or an immunologically reactive fragment thereof, and any combination thereof.
In a further embodiment, a kit of this invention can comprise, consist essentially of and/or consist of antigens (e.g., a diagnostic panel) and reagents for detecting, in a biological sample, the presence or absence of 1) an antibody that specifically reacts with Chlamydia trachomatis CT443 protein or an immunologically reactive fragment thereof, 2) an antibody that specifically reacts with Chlamydia trachomatis CT875 protein or an immunologically reactive fragment thereof, 3) Chlamydia trachomatis CT381 protein or an immunologically reactive fragment thereof and 4) an antigen of Chlamydia trachomatis CT147 protein or an immunologically reactive fragment thereof. Thus, a kit of this invention can include a diagnostic panel that comprises, consists essentially of or consists of an antigen of Chlamydia trachomatis CT443 protein or an immunologically reactive fragment thereof, an antigen of Chlamydia trachomatis CT875 protein or an immunologically reactive fragment thereof, an antigen of Chlamydia trachomatis CT381 protein or an immunologically reactive fragment thereof and an antigen of Chlamydia trachomatis CT147 protein or an immunologically reactive fragment thereof.
In another embodiment, a kit of this invention comprise, consist essentially of and/or consist of antigens (e.g., a diagnostic panel) and reagents for detecting, in a biological sample, the presence or absence of an antibody that specifically reacts with Chlamydia trachomatis CT875 protein or an immunologically reactive fragment thereof. Such a kit can further comprise, consist essentially of or consist of antigens (e.g., as additional components of a diagnostic panel) and reagents for detecting in a biological sample, the presence or absence of an antibody that specifically reacts with Chlamydia trachomatis CT147 protein or an immunologically reactive fragment thereof. Thus a kit of this invention can include a diagnostic panel that comprises, consists essentially of or consists of an antigen of Chlamydia trachomatis CT875 protein or an immunologically reactive fragment thereof. In some embodiments, such a kit can further comprise, consist essentially of or consist of Chlamydia trachomatis CT147 protein or an immunologically reactive fragment thereof.
It would be well understood by one of ordinary skill in the art that the kits of this invention can comprise one or more containers and/or receptacles to hold the reagents (e.g., antibodies, antigens, nucleic acids) of the kit, along with appropriate buffers and/or wash solutions and directions for using the kit, as would be well known in the art.
The following examples are included to demonstrate various embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventors to function well in the practice of the invention. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments that are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.

EXAMPLES

Example 1

Genome-Wide Identification of Chlamydia trachomatis Antigens Associated with Tubal Factor Infertility

Objective.
To identify C. trachomatis antigens that can be used to differentially diagnose tubal factor infertility in comparison to previously reported Heat Shock Protein 60 (HSP60).
Design.
In Vitro Study
Patients.
Infertile women with and without tubal pathology diagnosed laparoscopically.
Setting.
Academic medical center.
Main Outcome Measures.
Antibody responses to C. trachomatis in infertile women with or without tubal pathologies using a C. trachomatis genome-wide proteome array.
Results.
Comparison of the antibody profiles revealed 30 C. trachomatis antigens that were preferentially recognized by tubal factor infertility women with a detection sensitivity and specificity of 80.6% and 56.5%, respectively, 10 of which showed 100% specificity. A combination of CT443 and CT381 antigens yielded the highest detection sensitivity (67.7%) while maintaining 100% specificity.
Conclusion.
These findings have demonstrated that antibodies to CT443 and CT381, when used in combination, have higher sensitivity and specificity in predicting tubal factor infertility than other indicators for tubal factor infertility such as HSP60 antibodies (35.5%, 100%) or hysterosalpingogram (65%, 83%). Using a panel of C. trachomatis antigens to serologically diagnose tubal factor infertility can save the patients from undertaking expensive and invasive procedures for determining tubal pathology and choosing treatment plans.
Introduction.
Twenty-five to 35% of patients presenting for infertility evaluation have tubal disease (1-4). Chlamydia trachomatis is the primary sexually transmitted infection responsible for tubal factor infertility (TFI) (5-7) with C. trachomatis antibodies in approximately 70% of patients (8). C. trachomatis infected cells produce inflammatory cytokines (9-10) which may contribute to upper genital tract inflammatory damage (11-13). Lunefeld et al. found that among patients undergoing in vitro fertilization, those with C. trachomatis antibodies had decreased pregnancy rates (14).
C. trachomatis infection is often asymptomatic so patient history cannot dictate the presence of tubal disease (15-16). Elevated titers of anti-C. trachomatis antibodies are associated with TFI, but detection of overall antibody levels lacks the sensitivity and specificity required for differential diagnosis (17).
Measuring anti-C. trachomatis antibodies at the single antigen level may offer increased sensitivity and specificity for predicting TFI. Elevated anti-chlamydial heat shock protein 60 (HSP60, CT110) antibodies are associated with TFI (18-27). Anti-HSP60 antibodies are associated with decreased pregnancy rates in patients with an ectopic pregnancy history (17). When HSP60 antibodies are in follicular fluid, there are decreased implantation rates (28-29). Some have postulated that chlamydial HSP60 incites a strong inflammatory response that may cross-react with the highly conserved human HSP60 (25, 30-31). HSP60 may induce T-cell responses that contribute to the tubal damage (32-33).
In the present study a C. trachomatis whole-genome scale protein array has been developed that can profile antigen specificities of anti-Chlamydia trachomatis antibodies (34).
Human Patients.
Thirty one TFI and 23 IFC patients were enrolled at the University of Texas Health Science Center at San Antonio following Institutional Review Board approval. All women were at least 21 years old and underwent diagnostic laparoscopy with chromotubation as part of their infertility evaluation. Diagnosis of tubal infertility was defined as fallopian pathology consistent with hydrosalpinx, fimbrial phimosis, or peri-tubal adhesions. Exclusion criteria included prior tubal ligation, surgical finding of endometriosis, or a history of pelvic infection or inflammation other than pelvic inflammatory disease such as appendicitis. IFC patients had normal pelvic findings and tubal patency at laparoscopy. After the blood draw, serum samples were stored at −20° C. until analyzed.
Cell Culture and Chlamydial Infection.
As previously described, HeLa cells (American Type Culture Collection, Manassas Va. 20108) were cultured in DMEM (GIBCO PRL, Rockville, Md.) with 10% fetal calf serum (FCS; GIBCO BRL) at 37° C. with 5% carbon dioxide (CO₂) (34-36). C. trachomatis serovar D or Chlamydia pneumoniae AR39 organisms were grown, purified and titrated as previously described (36-38). For immunofluorescence assay, chlamydial organisms were used to infect HeLa cells grown on glass coverslips in 24-well plates. The sub-confluent HeLa cells were treated with DMEM containing 304 ml of DEAE-Dextran (Sigma, St. Louis, Mo.) for 10 minutes at 37° C. After removal of DEAE-Dextran solution, chlamydial organisms were added to the wells for 2 hours at 37° C. The infected cells were continuously cultured in DMEM with 10% FCS and 2 μg/ml of cycloheximide (Sigma, St. Louis, Mo.).
Immunofluorescence Assay (IFA).
Anti-chlamydial organism antibodies in human sera were titrated using an Immunofluorescence assay (IFA) as previously described (34, 36, 39, 40). Briefly, HeLa cells grown on coverslips were infected with C. trachomatis or C. pneumoniae organisms, fixed 48 h post-infection for C. trachomatis and 72 h for C. pneumoniae with 2% paraformaldehyde, and permeabilized with 2% saponin at room temperature for 1 hour. After blocking, human sera were added to the Chlamydia-infected cell samples. The primary Ab binding was visualized with a goat anti-human IgG conjugated with Cy3 (red; Jackson ImmunoResearch Laboratories, West Grove, Pa.), and DNA was labeled with Hoechst dye (blue; Sigma-Aldrich). The highest dilution of a serum that still gave a positive reactivity was defined as the titer of the given serum sample. Serum samples were serially diluted and the appropriate dilutions were repeated multiple times based on the results obtained from prior dilutions in order to obtain a more accurate titer for each serum. Images were acquired with an Olympus AX70 fluorescence microscope equipped with multiple filter sets (Olympus, Melville, N.Y.) as previously described (36, 40).
Chlamydial Fusion Protein-Arrayed Microplate Enzyme-Linked Immunosorbent Assay (ELISA).
Glutathione S-transferase (GST) fusion protein enzyme-linked immunosorbent assay (ELISA) for detecting human antibody recognition of chlamydial proteins was carried out as previously described (36). Bacterial lysates containing individual chlamydial GST fusion proteins were added to 96 well microplates pre-coated with glutathione (Pierce, Rockford, Ill.) at a 1:10 dilution in PBS with a total volume of 200 μl/well. Lysates containing GST alone, as negative, and GST-chlamydial protease-like activity factor (CPAF), as positive controls, were also included on each plate. The plates were incubated overnight at 4° C. to allow GST fusion proteins to bind to the plate-immobilized glutathione then blocked with 2.5% milk in PBS and washing with PBST (PBS with 0.05% Tween 20; Sigma Aldrich).
The human sera were pre-absorbed with bacterial lysates containing GST at 4° C. overnight, then incubated with glutathione beads (bioWorld, Dublin, Ohio) for 1 hour at room temperature to reduce background caused by non-specific human antibodies. The human antibody reactivity was detected with a goat anti-human-IgG, IgA and IgM conjugated with horse-radish peroxidase (HRP; Jackson ImmunoResearch Laboratories) plus the substrate 2,2′-azino-bi(2-ethylbenzothiazoline-6-sulforic acid)diammonium salt (ABTS; Sigma). The optical density (OD) was measured at 405 nm using a microplate reader (Molecular Devices Corporation, Sunnyvale, Calif.). To confirm the antibody binding specificity, all sera were further absorbed with lysates made from either HeLa cells alone or C. trachomatis serovar D-infected HeLa cells prior to reacting with the fusion protein-coated plates. The absorption was carried out as follows: HeLa cells with or without chlamydial infection were lysed via sonication at 2×10⁷cells per ml of PBS containing a cocktail of protease inhibitors. The pre-diluted serum samples were incubated with cell lysates overnight at 4° C. prior to reacting with the plate-immobilized chlamydial fusion proteins. The antibody binding that remained positive after HeLa-alone lysate absorption but significantly reduced by Chlamydia-HeLa lysate absorption was considered true positive.
Data Analyses.
Data were analyzed using SPSS v. 15.0 software (IBM, Chicago, Ill.) as previously described (36, 39). Briefly, titer values were log-transformed to produce a normal distribution and analyses were performed on transformed values. Student's t-Test was utilized to assess anti-C. trachomatis and anti-C. pneumoniae antibodies to evaluate overall mean differences between the 2 groups of patients. Because the a priori hypothesis was that the TFI group would have higher titers than the IFC group, a one-tailed analysis was used for the C. trachomatis data, but a two-tailed analysis was performed on the C. pneumonia data because there was no a priori hypothesis. Because the antibody titers had large variations within a given group, the serum titers were evaluated by ranges of <1:10 (Negative), 1:10 to 1:10,000 (Low), and >1:10,000 (High). Chi-Squared and Fisher's Exact Test were employed to compare overall antibodies to C. trachomatis and antibodies to C. pneumoniae.
ELISA results were analyzed using Student's t-Test and Fisher's Exact Test as appropriate. For the genome-wide ELISA, both Student's t-Test (for comparing quantitative OD value data) and Fisher's Exact Test (for comparing the number of sera positively reacted with a given antigen) were preformed. Using both methods allows for the identification of C. trachomatis antigens that are both clinically and statistically significant. When Student's t-Test was utilized, the OD values after subtracting background from the same plate were used. When Fisher's Exact Test was utilized, a response was determined positive when the OD value was equal to or greater than 2 standard deviations above the mean calculated from the same 96 well plate as described previously (39, 41).
Infertile Women with Laparoscopy-Identified Tubal Pathologies Developed Significantly Higher Titers of Anti-C. trachomatis Antibodies.
Sera from TFI or IFC were titrated using HeLa cells infected with either C. trachomatis or C. pneumoniae organisms as antigens in an IFA. TFI Patients developed high titers of antibodies to C. trachomatis (p<0.001) but not C, pneumoniae (p=0.269) (Table 1). When the patients were categorized based on levels of anti-chlamydial antibodies, most TFI patients developed high titers of anti-C. trachomatis antibodies (61.3%) while most IFC patients displayed lower titers (82.6%; p<0.001).
Table 1 shows titers of human antibodies against C. trachomatis and C. pneumoniae. Serum samples from women with TFI and IFC were 2 fold serially diluted starting with 1:10 and reacted with HeLa cells infected with either C. trachomatis or C. pneumoniae. The antibody reactivity was detected using an immunofluorescence assay as described herein. The highest dilution that still gave a positive reactivity was defined as the serum titer. Each serum sample was titrated in triplicate and the average was used as the geometric titer of a given serum sample. Student's t-Test was used to quantitatively analyze the differences between the two groups of patients. There is a statistically significant difference in titers of antibodies against C. trachomatis (p<0.001) but not C. pneumoniae (p=0.269) organisms. When the serum samples were divided into 3 categories (negative, low and high) based on antibody titers, the qualitative analysis with Chi-squared test still revealed a significant difference in the number of sera in different categories between the two groups of patients for antibodies against C. trachomatis (p<0.001) but not C. pneumoniae (p=0.634) organisms. Further pairwise Chi-squared analyses of the anti-C. trachomatis antibodies revealed significant differences between the high vs. low and high vs. negative groups. The number of individuals with high titers of anti-C. trachomatis antibodies in the TFI group is significantly higher than that in the IFC group.
Table 2 shows reactivity of 30 C. trachomatis antigens with 54 patient sera at 1:800 dilution. The 30 C. trachomatis antigens significantly recognized by 24 TFI patients were reacted with 54 patient sera (33 TFI and 21 IFC). All sera were diluted at 1:800 regardless of their overall anti-C, trachomatis antibody titers as determined with the immunofluorescence assay. The mean ODs of each antigen were compared between TFI and IFC groups using Student t-Test and the corresponding p values confirmed that all 30 antigens were significantly recognized by TFI patients. The number of positive recognition by either TFI or IFC was used to calculate recognition specificity and sensitivity as well as positive or negative predicting values (PPV or NPV; right panel). HSP60 (CT110) displayed a detection specificity of 82.6% and sensitivity of 71% and many other immunodominant antigens such as pCTO3 (Pgp3, a plasmid-encoded secreted protein), CT858 (CPAF, a chlamydial protease/proteasome-like activity factor that is secreted into host cell cytosol), CT823 (cHtrA, a secreted stress response serine protease), CT813 (an inclusion membrane protein), CT443 (OmcB, outer membrane complex protein B) and CT143 (a hypothetical protein) behaved similarly. Only the hypothetical protein CT557 had 100% specificity but its sensitivity was only 29%. Thus, under this assay condition, no single antigen or combinations of antigens can achieve 100% specificity with a sensitivity of >50%.
Identification of C. trachomatis Antigens Preferentially Recognized by Infertile Women with or without Tubal Pathology.
It is difficult to use the quantitative difference in overall anti-C. trachomatis antibodies to diagnose TFI. To identify antigens that are recognized by TFI patients, anti-C. trachomatis antibodies in 24 TFI and 11 IFC patients were mapped at the genome-wide scale since these patients displayed an overall anti-C. trachomatis antibody titer above 1:1000. These 35 sera recognized C. trachomatis antigens distributed across the genome with 265 antigens recognized by at least one antiserum and 643 antigens not detected by any sera. Many C. trachomatis antigens are recognized by both groups of patients, but there are antigens preferentially recognized by either group. Thirty antigens were significantly recognized by TFI based on either mean OD values (Student's t-test) or recognition frequency (Fisher's exact test). Reactivity was confirmed using absorption against either HeLa alone or C. trachomatis-infected HeLa lysates as described previously (36).
Identification of C. trachomatis Antigens Uniquely Recognized by TFI Patients.
To identify antigens that can be used to predict TFI in infertility clinics, antigens were identified that were uniquely recognized by TFI patients. The 30 antigens preferentially recognized by TFI patients (Table 2) were reacted with sera from all 54 patients (including 31 TFI and 23 IFC) regardless of their overall anti-C. trachomatis antibody titers, The HSP60 (CT110) reacted with 22 of the 31 TFI and 4 of the 23 IFC sera with a specificity and sensitivity of 82.6% and 71.0%, respectively, in predicting TFI.
To further increase specificity, a 5-fold dilution (final dilution of 1:4000) was used to reduce the false-positive rate. At this dilution, 10 of the 30 antigens, including CT110, CT322, CT376, CT381, CT414, CT443, CT681, CT795, CT798 and CT813, failed to react with any sera from the IFC group. Thus, these 10 antigens were uniquely recognized by TFI patients with a detection specificity of 100%. Dilution of the samples decreased detection sensitivity. HSP60 (CT110) only reacted with 11 out of 31 TFI sera, dropping the detection sensitivity to 35.5% along with three immunodominant antigens (CT795, CT798, CT813). CT443 reacted with 18 of the 31 TFI sera, maintaining a sensitivity of 58.1%. When the reactivity of the 10 antigens was analyzed at each individual antiserum level, it was found that the 10 antigens all together reacted with 21 independent sera of 31 total TFI sera, with a sensitivity of 67.7%. More importantly, this sensitivity can be maintained using fewer antigens. Combining HSP60 with CT376, CT381 and CT798 (total 4 antigens) or CT443 with CT381 (only 2 antigens) maintains sensitivity of 67.7%. As for other immunodominant antigens with a detection specificity of <100%, their detection sensitivity can be 80% even after serum dilution. Due to their ability to generate false positive results by reacting with IFC samples, it is clinically undesirable to use these antigens for screening for TFI.
C. trachomatis organisms cause pathologies in the fallopian tubes, leading to complications such as ectopic pregnancy and infertility. Since infertility can be caused by many different factors, distinguishing tubal infertility from other causes is useful for guiding treatment plans. The goal of the current study is to identify C. trachomatis antigens that can improve specificity and sensitivity in detecting TFI.
The finding that anti-C. trachomatis but not anti-C. pneumoniae antibodies are highly associated with TFI is consistent with the literature (42-44). Efforts have been made to develop individual C. trachomatis antigen-based detection methods. Previous reports demonstrate that anti-HSP60 antibodies are detected in 70-80% of TFI patients (21-22, 45-46). The present genome-wide search for additional markers of TFI not only confirmed these findings but also revealed new information for further increasing specificity and sensitivity in detecting TFI.
Thirty antigens were preferentially recognized by TFI patients. At 1:800 dilution, HSP60 reacted with 22 of the 31 TFI sera (71% sensitivity) and 4 of the 23 IFC sera (82.6% specificity), which is consistent with previous findings and suggests that the ELISA described herein is comparable to other assays. When the sera were diluted to 1:4000 in order to further increase specificity, 10 of the 30 antigens achieved 100% specificity. Although the sensitivity decreased, careful examination revealed that a combination of 2 antigens [CT381 and CT443 (Outer membrane complex B, OmcB)], or 4 antigens [CT110 (HSP60), CT376, CT381 & CT798] detected TFI with a specificity and sensitivity of 100% and 67.7%, respectively. These combinations of antigens improved the C. trachomatis serology approach for diagnosing tubal infertility over using HSP60 alone (35.5% sensitivity), which represents a clinically significant improvement.
Hysterosalpingogram (HSG) has a detection specificity and sensitivity of 83% and 65% respectively for detecting tubal pathology (47). C. trachomatis antigen-based serology diagnosis has numerous advantages over HSG besides improved detection, including sparing patients from the discomfort, radiation, and potential for infectious sequellae. This conclusion is consistent with previous reports that elevated chlamydial antibody levels are comparable to HSG (48) in diagnosing TFI and that HSG does not add to the medical knowledge on whether C. trachomatis infection contribute to tubal pathology (49).
It is unknown whether these antigens themselves or antigen-specific immune responses contribute to the inflammatory pathologies in the fallopian tubes. The protein CT443, or OmcB, displayed the highest rate of reactivity with TFI patient sera. OmcB is a highly conserved immunodominant antigen, but the precise location of OmcB in the organisms and its role during infection is poorly understood (34, 50-52).
Despite the overwhelming evidence of C. trachomatis infection association with TFI, not every patient is infected with C. trachomatis or developed immune responses to C. trachomatis. Interventions such as early antibiotic therapy may cause a negative or low titer in patients, but it is unlikely that tubal pathology would be attributed to the C. trachomatis infection in these patients. Tubal pathology in TFI patients without positive C. trachomatis titers might be caused by other sources of infection such as Neisseria gonorrhoeae (53-54) and Mycoplasma genitalium (55-56). Thus, to further increase the sensitivity in diagnosing TFI, other infection causes should also be taken into account.

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Example 2

Chlamydia trachomatis Antigens Recognized by Women with Tubal Factor Infertility, Normal Fertility and Acute Infection

Abstract.

To identify C, trachomatis antigens associated with tubal factor infertility, a whole genome scale C. trachomatis proteome array was used to compare antibody specificity profiles among women with tubal factor infertility, normal fertility and acute C. trachomatis infection. Thirteen immunodominant antigens reacted with ≧50% antisera from all women. Ten C. trachomatis antigens were uniquely recognized by women diagnosed with tubal factor infertility. Assessing antigen fragments with serum sample dilution, chlamydial antigens HSP60, CT376, CT557 & CT443 could discriminate between women with tubal factor infertility and women with normal fertility at a sensitivity of 63% and specificity of 100%, respectively. However, these antigens also strongly reacted with antisera from women diagnosed with acute C. trachomatis infections. Nevertheless, women with an acute infection preferentially recognized a new set of 21 antigens. Of these, CT875 & CT147 together distinguished women with acute infection from all other C. trachomatis-exposed women with a detection sensitivity of 63% and specificity of 100%, respectively. A combination of both sets of antigens may be useful to screen all women for tubal factor infertility and identify acute C. trachomatis infection.
Introduction.
Chlamydia trachomatis is a leading cause of sexually transmitted bacterial infection (STI) in the US, affecting over one million women in 2008 alone (1). The incidence of C. trachomatis infection has steadily increased since it was first recorded in 1984, and continues to rise yearly. C. trachomatis infection is often undiagnosed and untreated because it is asymptomatic. If untreated, C. trachomatis infection may lead to ascending infection, causing complications such as ectopic pregnancy and tubal factor infertility (1-2). However, not all women infected with C. trachomatis develop tubal damage. Up to 50% of women with normal fallopian tubes on laparoscopy or normal fertility have developed high titers of anti-C. trachomatis antibodies (3). It is not clear what determines whether a C. trachomatis-exposed woman will develop tubal pathology and complications. Nevertheless, the severity and number of episodes of C. trachomatis infection can contribute to tubal pathology. Since host immune responses reflect both infection status and host responsiveness to infection, monitoring specific antibody responses to C. trachomatis infection may lead to the identification of biomarkers that correlate with tubal pathology. Women with robust antibody responses to the C. trachomatis major outer membrane protein (MOMP) are less likely to develop tubal pathology while those who generate high titers of anti-HSP60 antibodies are at a significantly higher risk of developing tubal factor infertility (TFI) (4). High resolution mapping of host antibody responses to C. trachomatis infection may aid in the identification of C. trachomatis antigens associated with TFI.
Previous studies have revealed a strong association of anti-C. trachomatis and anti-HSP60 antibodies with TFI (3-9). Using a high-resolution whole genome scale protein array assay, the profiles of antigen specificities of anti-C, trachomatis antibodies were compared between women with TFI and women with infertility due to other causes (infertility control, IFC). The association of HSP60 antibodies with TFI was confirmed and additional new C. trachomatis antigens associated with TFI were identified (5). However, these and other studies focused on well-defined study subjects for comparison. When more diverse patient populations were included in the comparison, the anti-C. trachomatis and HSP60 antibodies no longer significantly correlated with tubal factor infertility or subfertility (10). The goal of the current study is to identify C. trachomatis antigens that are preferentially recognized by different groups of women with diverse socio-economic status. It was found that C. trachomatis antigens such as HSP60, which have previously been shown to be associated with TFI (5), were also highly reactive with antisera from women with acute C. trachomatis urogenital tract infection. Thus, TFI-associated antigens including HSP60 cannot predict tubal factor infertility in the general female population. Here, a whole genome scale proteome array was used to define a set of 21 antigens for distinguishing women with acute C. trachomatis infection from other C. trachomatis-exposed women. It is demonstrated that distinct panels of C. trachomatis antigens may be used to predict pathology and/or clinic phenotypes caused by C. trachomatis infection.
Human Subjects.
All patients were enrolled at the University of Texas Health Science Center at San Antonio after receipt of Institutional Review Board (IRB) approval. All participants underwent a single blood draw. Serum samples were stored at −20° C. until analyzed. Subjects were composed of three distinct socioeconomic and demographic groups. The TFI group (N=24) was recruited from a university-based infertility clinic. The typical payer mix is 13% privately insured, 69% self pay, and 18% military. Subjects with TFI were eligible if they were at least 21 years of age and had a pelvic laparoscopy demonstrating hydrosalpinx, fimbrial phimosis, or peritubal adhesions. Exclusion criteria included prior tubal ligation, surgical finding of endometriosis, or a history of pelvic infection or inflammation other than pelvic inflammatory disease, such as appendicitis. The fertile control (FC) group (N=25) was recruited from a university affiliated county hospital. The typical payer mix is 31% privately insured, 29% Medicare, 36% Medicaid. FC subjects were eligible if they were at least 21 years of age and had at least one live birth and normal pelvic findings at the time of tubal ligation. FC patients were enrolled at the time of their scheduled tubal ligation. Sterilization was performed either via laparoscopy or via mini-laparotomy. The sexually transmitted infection (STI) group (N=24) was referred by the health department to a university based STI clinic after diagnosis of any sexually transmitted infections or diseases. Patients reported their insurance as 11% privately insured, 25% Medicaid, 25% uninsured and 14% did not respond. STI patients were eligible if they were of reproductive age (15-45 years of age) and had a recent diagnosis of C. trachomatis infection. The diagnosis was based on positive C. trachomatis nucleic acid detection in vaginal swab samples as described previously (11-12). Blood was taken within 2 weeks of the diagnosis.
Cell Culture and Chlamydial Infection.
As previously described (13-14), HeLa cells (American Type Culture Collection) were cultured in Dulbecco's minimum essential medium (DMEM; GIBCO) with 10% fetal calf serum (GIBCO) at 37° C. with 5% CO₂ . C. trachomatis serovar D or C. pneumoniae AR39 organisms were grown, purified, and titrated as previously described (3, 5, 13, 15). For immunofluorescence assay, chlamydial organisms were used to infect HeLa cells grown on glass coverslips in 24-well plates. The subconfluent HeLa cells were treated with DMEM containing 30 mg/mL of DEAE-Dextran (Sigma) for 10 minutes at 37° C. After removal of DEAE-Dextran solution, chlamydial organisms were added to the wells for 2 hours at 37° C. The infected cells were continuously cultured in DMEM with 10% fetal calf serum and 2 mg/mL of cycloheximide (Sigma) for 48 h (C. trachomatis) or 72 h (C. pneumoniae) before sample processing.
Immunofluorescence Assay.
Antichlamydial organism antibodies in human sera were titrated using an immunofluorescence assay as previously described (3, 5). Briefly, HeLa cells grown on coverslips were infected with C. trachomatis or C. pneumoniae organisms, fixed with 2% paraformaldehyde, and permeabilized with 2% saponin at room temperature for 1 hour. After blocking, human sera were added to the Chlamydia-infected cell samples. The primary antibody binding was visualized with a goat anti-human IgG conjugated with Cy3 (red; Jackson ImmunoResearch Laboratories), and DNA was labeled with Hoechst dye (blue; Sigma-Aldrich). The highest dilution of a serum that still gave a positive reactivity was defined as the titer of the given serum sample. Serum samples were serially diluted, and the appropriate dilutions were repeated multiple times according to the results obtained from prior dilutions, to obtain a more accurate titer for each serum. Images were acquired with an Olympus AX70 fluorescence microscope equipped with multiple filter sets, as previously described (16-17).
Chlamydial Fusion Protein-Arrayed Microplate ELISA.
Glutathione S-transferase (GST) fusion protein ELISA for detecting human antibody recognition of chlamydial proteins was carried out as previously described (11-12). Bacterial lysates containing individual chlamydial GST fusion proteins were added to 96-well microplates precoated with glutathione (Pierce) at a 1:10 dilution in PBS, with a total volume of 200 μL per well. Lysates containing GST alone, as negative, and GST-chlamydial protease/proteasome-like activity factor (CPAF), as positive controls, were also included on each plate. The plates were incubated overnight at 4° C. to allow GST fusion proteins to bind to the plate-immobilized glutathione, then blocked with 2.5% milk in phosphate-buffered saline (PBS) and washed with PBST (PBS with 0.05% Tween 20; Sigma-Aldrich). The human sera were preabsorbed with a bacterial lysate containing GST at 4° C. overnight, then incubated with glutathione beads (bioWorld) for 1 hour at room temperature to reduce background caused by nonspecific human antibodies. The human antibody reactivity was detected with a goat anti-human IgG, IgA, and IgM conjugated with horseradish peroxidase (Jackson ImmunoResearch Laboratories) plus the substrate 2,20-azino-bi (2-ethylbenzothiazoline-6-sulforic acid)diammonium salt (Sigma). The optical density (OD) was measured at 405 nm using a microplate reader (Molecular Devices). To confirm the antibody binding specificity, all sera were further absorbed with lysates made from either HeLa cells alone or C. trachomatis serovar D-infected HeLa cells before reacting with the fusion protein-coated plates. The absorption was carried out as follows: HeLa cells with or without chlamydial infection were lysed via sonication at 2×10⁷cells per milliliter of PBS containing a cocktail of protease inhibitors. The prediluted serum samples were incubated with cell lysates overnight at 4° C. before reacting with the plate-immobilized chlamydial fusion proteins. The antibody binding that remained positive after HeLa-alone lysate absorption but significantly reduced by Chlamydia-HeLa lysate absorption was considered true positive.
Data Analyses.
Data were analyzed using Microsoft Excel 2007. ANOVA was used to compare anti-C. trachomatis and anti-C. pneumoniae antibody titers between all three groups of patients. Student's t-Test was used to identify statistical differences between given two groups. Results from ELISA were analyzed using both Student's t-Test (for comparing quantitative OD value data) and Chi-squared or Fisher's Exact test (for comparing the number of sera positively reacted with a given antigen). Combination of these two methods allowed for the identification of C. trachomatis antigens that are both clinically and statistically significant. When Student's t-Test was used, the OD values after subtracting background from the same plate were used. When the Chi squared test was used, positive reactivity frequency was used. A reaction was determined positive when the OD value was 2 standard deviations (SD) above the mean calculated from the same 96-well plate (5).
Overall Anti-C. trachomatis and Anti-C. pneumoniae Antibodies in Three Groups of Women.
Serially diluted serum samples from women with tubal factor infertility (TFI), normal fertility (FC) and acute C. trachomatis infection (STI) were reacted with C. trachomatis and C. pneumoniae-infected HeLa cells to measure the titers of the corresponding antibodies (Table 3). STI women displayed the lowest levels of anti-C. pneumoniae antibodies compared to TFI and FC groups (P<0.05 for both, Student's t-Test) while TFI and FC displayed similar levels of the antibodies (p=0.34). The distribution of anti-C. pneumoniae antibodies correlated well with the age distribution among the three groups of women. STI women were significantly younger (mean age 21.8±3.1 years) than both TFI (34.6±4.4, p<0.05) and FC (32.5±5.7, p<0.05), respectively, while the TFI and FC women displayed similar ages (p>0.05). These results are consistent with a general concept that anti-C. pneumoniae antibody levels increase as age increases (18-19).
However, the anti-C. trachomatis antibody titers were highest in the STI group compared to those in either the TFI or FC groups (p=0.0748 & p=0.0099, respectively) while there was no significant difference between TFI and FC groups (p=0.2229). The STI group was significantly younger than both the TFI and FC groups as noted above. Thus, in contrast to the age-dependent increase in anti-C. pneumoniae antibody titers, the anti-C. trachomatis antibody titers decreased as age progressed, probably due to reduced exposure to C. trachomatis. This observation is consistent with CDC data showing that adolescents and young women ages 15 to 24 are at the highest risk for STIs (2). Although the titer of anti-C. trachomatis antibodies in TFI women (35483.33±39950.49) appeared to be higher than that in FC women (23760.00±23974.4), the difference was not statistically significant (p=0.2229), which stands in contrast to the previous report that women with tubal factor infertility displayed significantly higher levels of anti-C. trachomatis antibodies than women with infertility due to non-tubal causes (3, 5). The previous studies focused on infertile women recruited from an infertility clinic while the above analysis compared tubal factor infertility women from the infertility clinic with women of normal fertility. Clearly, when diverse female populations are included, the overall anti-C. trachomatis antibody levels are no longer associated with tubal factor infertility. A whole genome scale protein array was used next to profile antigen-specificities of antibodies in the serum samples from this diverse population of women.
Genome-Wide Antibody Profiles Reveal Immunodominant Antigens Commonly Recognized by all Three Groups of Women.
Anti-C. trachomatis antibodies from 24 TFI, 25 FC and 24 STI women were profiled for their antigen-specificities at a genome-wide scale. These 73 antisera recognized C. trachomatis antigens distributed across the entire genome with 541 antigens recognized by at least one antiserum and 367 antigens not detected by any antisera. Many C. trachomatis antigens were recognized by all 3 groups of women although some antigens were preferentially recognized by individual or combinations of groups. Regardless of which groups the women were from, 50% or more of the 73 women recognized a total of 13 antigens, including pCT03 (93% frequency) (Pgp3, a plasmid-encoded hypothetical protein that is secreted into host cell cytosol (20-21)), CT858 (90% frequency) (CPAF, a chlamydial protease/proteasome-like activity factor known to be secreted into host cell cytosol (14)), CT101 (79% frequency) (hypothetical protein, HP), CT841 (77% frequency) (FtsH, ATP-dependent Zinc protease), CT240 (73% frequency) (Recombination protein RecR), CT443 (73% frequency) (outer membrane complex protein B, OmcB (22)), CT142 (60% frequency) and CT143 (71% frequency) (both HPs), CT813 (68% frequency) and CT529 (62% frequency) (both inclusion membrane proteins, Incs (23, 24)), CT694 (66% frequency) (a putative effector of the type III secretion pathway (25)), CT022 (55% frequency) (50S ribosomal protein L31 type B) and CT806 (55% frequency) (insulinase family/protease III, Ptr). These proteins are designated as immunodominant antigens in these women.
Identification of Antigens Preferentially Recognized by TFI Women.
The reactivity of the antiserum samples with each of the 908 C. trachomatis antigens was compared between the 3 groups of women both quantitatively (comparison of OD values using ANOVA) and qualitatively (comparison of recognition frequency using Fisher's Exact test). Antigens that displayed statistically significant differences in antibody reactivity (either quantitatively or qualitatively) and were recognized by 10% or more of the antisera from at least one group of women were selected. There were a total of 97 antigens that met both the above requirements. The use of 10% cutoff for recognition frequency was set to eliminate antigens with extremely low reactivities that were no longer biologically significant, although statistically significant. Many antigens shown at the bottom half of the figure were visually identifiable as antigens preferentially recognized by women from one or two groups but not all groups. Since a major goal of the current study is to identify TFI-associated antigens, the antigens that displayed statistically significant differences in recognition by TFI versus FC women were analyzed first (Table 4). Among the 10 antigens that were preferentially recognized by TFI women, 5 failed to react with any antisera from FC women under the current serum dilution (1:1000), suggesting that these antigens can distinguish TFI from FC women with 100% specificity. When the reactivity patterns of these 5 antigens with TFI women were revealed (FIG. 2), it was found that CT110, the chlamydial HSP60, reacted with antisera from 9 out of 24 TFI women, thus with a detection sensitivity of 38%. Antigen CT376 reacted with additional 3 TFI samples while CT557 reacted with another 2 TFI samples. Together, these 3 antigens detected TFI samples with a sensitivity of 58% while still maintaining 100% specificity. Among the remaining 5 antigens recognized by both TFI and FC women, CT443, the outer membrane complex protein B (OmcB) was recognized by TFI women with the highest frequency and intensity (both were significantly higher compared to FC women). Testing was done to determine whether a combination of human serum dilution and CT443 fragments could help to further differentiate TFI from FC women (FIG. 3). The CT443 full length (FL) proteins were expressed in 12 different fragments, all of which were used to react with both TFI and FC women antisera at different dilutions. Only the fragments representing the C-terminal portion of CT443, including fragments 9 (F9), 11 and 12, were reactive with human antisera. Nevertheless, at each human serum dilution, the CT443 fragments produced significantly greater differences than the full length CT443 did in antibody reactivity between TFI and FC samples. For example, at 1:1000 dilution of human sera, the full length CT443 reacted with 88% of TFI and 60% of FC women (a 32% decrease in reactivity with FC women antibodies) while F9 reacted with 42% TFI and only 8% FC (81% decrease), F11 reacted with 58% TFI and 24% FC (59% decrease) and F12 reacted with 50% TFI and 16% FC (48% decrease). When the human antiserum was used at 1:3000 dilution, F11 still reacted with 50% TFI but only 4% FC, representing a 92% decrease in reactivity with FC. Only one of the 25 FC antisera remained reactive with F11. Further modification of F11 may lead to the elimination of the FC reactivity while maintaining high levels of reactivity with TFI. These results have provided proof of principle that fragmentation of immunodominant cross-reactive antigens may represent a viable approach for identifying antigens with increasing differential diagnosis power. Interestingly, when human sera were diluted to 1:10,000, the full length CT443 no longer reacted with any FC women samples but retained reactivity with 38% of TFI samples, suggesting that serum sample dilution may represent another strategy to increase the ability to differentiate between women with different phenotypes. When the reactivity patterns of CT443 and its fragment 11 were compared with those of HSP60, CT376 and CT557, one additional TFI antiserum was detected (FIGS. 4A-B). Thus, a combination of these 4 antigens achieved a detection sensitivity of 63% for TFI women while still maintaining 100% detection specificity when both TFI and FC women were included.
Identification of Antigens Preferentially Recognized by Women with Acute C. trachomatis Infection.
When the antigens that were demonstrated to be able to discriminate TFI from FC samples with 100% specificity were applied to women diagnosed with acute C. trachomatis infection from a STI clinic, these antigens also strongly reacted with the STI serum samples even when the samples were highly diluted (FIG. 4). This is because STI women were all diagnosed with acute C. trachomatis infection when sera were collected and they developed high titers of antibodies to many antigens as revealed in Table 3. Fortunately, the whole genome scale proteome array also allowed the identification of C. trachomatis antigens that were preferentially recognized by STI but not TFI women (Table 5). A total of 21 antigens that displayed the greatest differences in reactivity between STI and TFI women were selected based on both quantitative and qualitative statistical analyses. It appeared that all 21 antigens strongly reacted with samples from patients with STIs, but were poorly recognized by either TFI or FC women serum samples. For example, when human serum samples were diluted at 1:1000, CT147 reacted with 88% of STI, but only 21% of TFI or 16% of FC samples and CT875 was recognized by 75% STI women, but only 17% of TFI or 16% FC women. Clearly, these 21 antigens were highly associated with acute infection since only the STI women were detected positive for C. trachomatis DNA in vaginal swab samples at the time when sera were taken while neither TFI nor FC women displayed any sign of acute C. trachomatis infection. These antigens could be used to distinguish STI patients from those in other groups. These antigens have been designated as acute infection-associated antigens. When human serum samples were diluted at 1:10,000, 13 of the 21 acute infection-associated antigens maintained positive reactivity with STI samples, but failed to react with any serum samples from either TFI or FC women (FIG. 5). The reactivity patterns of these 13 acute infection-associated antigens revealed that CT875 reacted with 14 out of the 24 STI serum samples. CT147 reacted with an additional STI sample (although the rest reacted with STI samples already detected by CT875 and CT147). Thus, antigens CT875 and CT147 together can be used to distinguish STI women from the rest of women with a detection sensitivity of 63% and a specificity of 100%. Continuing optimization of the detection system and use of acute infection-associated antigen fragments may allow for further increases in detection sensitivity while maintaining high specificity.
Table 3 shows titers of antibodies against C. trachomatis and C. pneumoniae in women with tubal factor infertility (TFI), normal fertility (FC) or acute infection (STI). Serum samples from women with TFI, FC, or STI were 2-fold serially diluted starting with 1:10 and reacted with HeLa cells infected with either C trachomatis or C pneumoniae. The highest dilution that still gave a positive reactivity was defined as the serum titer. ANOVA was used to analyze the overall differences among the 3 groups. There is a statistically significant difference in titers of antibodies against both C. trachomatis and C. pneumoniae. However, further pairwise analyses with Student's t-Test reveals significantly higher titers of anti-C. trachomatis but lower titers of anti-C. pneumoniae antibodies in STI women.
Table 4 shows antigens preferentially recognized by women with tubal factor infertility (TFI) but not normal fertility (FC). When Fisher's Exact test and Student's t-Test were used to compare each of 908 protein reactivities between TFI and FC groups, 10 antigens displayed statistically significantly differences by either method. Their corresponding ORF names, reactivity frequency (Freq) and intensity (OD values: mean plus/minus standard deviations) along with the p values were listed in corresponding columns. Five of the 10 antigens (CT110, CT376, CT111, CT557 & CT579) reacted with no FC samples.
Table 5 shows antigens that are most significantly recognized by women with acute C. trachomatis infection. Each of the 541 human antibody-reactive C. trachomatis antigens was compared for their reactivities with antisera from STI versus TFI or FC women using Student's t-Test (for comparing OD values in mean plus/minus standard deviations) and Fisher's Exact Test (for comparing reactive frequencies, Freq; a positive reaction was identified if the OD value is equal to or above the mean plus 2 standard deviations). Antigens that displayed the most differences and were recognized by STI women with a recognition frequency of 30% or higher were selected. A total of 21 antigens met the two requirements and were listed from top to bottom based on the extent of differential recognition by STI versus TFI or FC women. The reactivity was measured using a human serum dilution of 1:1000. All antigens were highly reactive with STI women samples and significantly less reactive with both TFI and FC samples.
In this study, the whole genome scale proteome array analyses have been extended to women with TFI, normal fertility or acute infection from a diverse socioeconomic group of women in San Antonio, Tex. The overall goal is to define C. trachomatis antigens that can be used to differentiate women with different clinical phenotypes after C. trachomatis infection with a focus on identifying biomarkers for predicting/diagnosing tubal factor infertility. The 3 groups of women recruited into the current study can largely represent the general female population since their overall anti-C. pneumoniae antibody levels increase as ages progress as demonstrated in other populations of women (18-19) while the overall anti-C. trachomatis antibody levels are the highest in STI women with an age range that is known to be most susceptible to C. trachomatis infection in the US (2). Although the 3 groups of women recognized many common antigens, quantitative and qualitative statistical analyses have allowed for the identification of 10 antigens that were preferentially recognized by TFI but not FC groups. Using serum sample dilution and antigen fragmentation to increase differentiating capacity, a 4-antigen combination consisting of HSP60, CT376, CT557 and CT443 distinguished tubal factor infertility from normal fertility women with a detection sensitivity of 63% and specificity of 100%. However, these same antigens also strongly reacted with antisera from women diagnosed with acute C. trachomatis from a STI clinic. Fortunately, the whole genome proteome array has led to the identification of 21 antigens that are highly associated with acute infection, two of which, CT875 & CT147, distinguished women with acute infection from other C. trachomatis-exposed women with a detection sensitivity of 63% and specificity of 100%. Thus, it is possible to use a combination of antigens to screen all women for predicting/diagnosing chlamydial infection and diseases.
Many previous studies have shown an association of anti-C. trachomatis antibodies with tubal factor infertility or subfertility (3, 5-7). Many of these studies focused on the comparison between women with tubal factor infertility and infertile women due to non-tubal causes but both visiting the same infertility clinics. However, when more diverse female populations were analyzed and included in the comparison, the anti-C. trachomatis antibodies were no longer significantly higher in patients with tubal factor infertility (10). In the current study, no statistically significant difference in anti-C. trachomatis antibody titers was found between women diagnosed with tubal factor infertility from a infertility clinic and women with normal fertility undergoing tubal ligation procedures. Thus, the overall anti-C. trachomatis antibodies, although highly associated with tubal factor infertility when compared to infertile women, cannot predict tubal factor infertility among C. trachomatis-exposed women.
It has been previously shown that chlamydial HSP60 detected tubal factor infertility with a sensitivity of 36% and a specificity of 100% while a combination of antigens CT443 and CT381 provided a detection sensitivity of 68% while still maintaining 100% specificity (5). In the current study, when the antigen specificity profiles of anti-C. trachomatis antibodies in the TFI women were compared with those in FC women undergoing tubal ligation, these previous overall findings still held true. The chlamydial HSP60 along with antigens CT376 & CT557 positively detected 14 out of the 24 TFI samples with a sensitivity of 58%, but failed to react with any serum samples from FC women (100% specificity) (FIG. 6). CT443 or OmcB (outer membrane complex protein B) was found to react with serum samples only from tubal factor but not non-tubal infertility patients in the previous study (5). Although CT443 was recognized by both TFI and FC women in the current study, the reactivity of CT443 with TFI women was significantly stronger than that with FC women.
Using antigen fragmentation and serum dilution, it was possible to increase specificity of this diagnosis. By increasing the serum dilution, CT443 or CT443 fragments were able to differentiate TFI from FC samples. Using a dilution of 1:3,000, CT443 F11 reacted with 50% TFI but only 4% FC. Additional modifications of F11 may eliminate the residual FC reactivity. Further dilution of human sera to 1:10,000 completely removed the reactivity of CT443 with FC samples but allowed reactivity of CT443 with 38% of TFI samples. Together, these observations have provided a proof of principle that serum dilution and fragmentation of immunodominant cross-reactive antigens may represent viable approaches for identifying biomarkers to diagnose pathologies or predict clinic phenotypes associated with C. trachomatis.
Although the antigens identified above can distinguish TFI from FC women with 100% specificity, these antigens were also found to be strongly reactive with serum samples from women diagnosed with acute C. trachomatis infection from a STI clinic. Fortunately, the power of the whole genome scale proteome array has also allowed or the identification of 21 C. trachomatis antigens that were preferentially recognized by STI but not TFI nor FC women. Two of the 21 acute infection-associated antigens, CT875 and CT147, distinguished STI women from TFI and FC women with a detection sensitivity of 63% and a specificity of 100%. Continued optimization of the detection system and use of acute infection-associated antigen fragments may allow for further increases in detection sensitivity while maintaining high specificities. The observation that the acute infection-associated antigens were all poorly recognized by both TFI and FC women suggests that women in the TFI and FC groups were exposed to C. trachomatis in the past without ongoing infection. All STI women were detected positive for C. trachomatis DNA in vaginal swab samples at the time when sera were taken (11-12). However, the C. trachomatis status in either TFI or FC women was not specifically monitored when blood samples were collected. Nevertheless, physical examination revealed that none of the TFI and FC women displayed any sign of acute C. trachomatis infection in the lower genital tract (data not shown) and the nucleic acid detection rate was very low in patients who visited the same infertility or OB/GYN clinics in the past. Thus, the acute infection-associated antigens may represent useful biomarkers for indicating recent infection.
Identification of biomarkers associated with pathologies and/or clinic phenotypes caused by C. trachomatis infection will not only provide safer means for diagnosis or prognosis of C. trachomatis infection and diseases but may also promote better understanding of the mechanisms of C. trachomatis pathogenesis. It is widely believed that the association of chlamydial HSP60 with TFI suggests a role of HSP60 or host immune responses to HSP60 in tubal pathologies. Due to the high degree of amino acid sequence homology between chlamydial and human HSP60, anti-chlamydial HSP60 antibodies may cross-react with host HSP60 (26-27) or anti-HSP60 T cell responses may attack fallopian tube tissues (28-29). However, a recent study has shown that although antibodies reactive with chlamydial HSP60 were significantly higher in TFI women than those in control group, antibodies reactive with human HSP60 were at similar levels in both groups (30), suggesting that the anti-HSP60 cross-reactive antibodies may not significantly contribute to chlamydial pathogenesis. Chlamydial HSP60 has been shown to activate inflammatory pathways in macrophages (31), which may allow HSP60 to exacerbate inflammatory damage in fallopian tubes. The other C. trachomatis antigens that are significantly associated with TFI including CT376, CT557 & CT443 may also contribute to tubal pathology. Both CT376 and CT557 are highly conserved metabolic enzymes: malate dehydrogenase (334 amino acids) and dihydrolipoamide dehydrogenase (pyruvate dehydrogenase E3 component, 465 amino acids), respectively. These enzymes share high degrees of amino acid sequence homology with their host counterparts with 43% for CT376 and 38% for CT557, respectively. These metabolic enzymes are supposed to be localized in the cytoplasm of chlamydial organisms and should not be leaked into host cells during chlamydial infection. As with the cytoplasmic HSP60, CT376 and CT557 may be released into host environments after RB lysis. Clearly, both CT376 and CT557 are exposed to human immune cells during chlamydial infection in humans since humans developed robust antibody responses to these proteins. The question is whether CT376 and CT557 can be as inflammatory as HSP60 and immune responses to these proteins can contribute to chlamydial pathogenesis. CT443 or OmcB is an immunodominant protein in the outer membrane complex known as outer membrane complex protein B, although its precise location in chlamydial organisms remains unclear. It has been shown that a CT443 C-terminal fragment is released into host cell cytosol (22), which may explain the high immunogenicity of CT443 C-terminus since chlamydial proteins secreted into or exposed to host cell cytosol are known to be more immunogenic (32-33). Consistent with this concept is the current finding that most human antibodies recognized CT443 fragments covering the C-terminal regions and a previous report that a CD8+ T cell epitope was mapped to CT443 C-terminus (34).

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20. Li et al. The chlamydial plasmid-encoded protein pgp3 is secreted into the cytosol of Chlamydia-infected cells. Infect Immun 2008 August; 76(8):3415-28.
21. Chen et al. Characterization of Pgp3, a Chlamydia trachomatis plasmid-encoded immunodominant antigen. J Bacteriol 2010 Sep. 17.
22. Qi et al. A Chlamydia trachomatis OmcB C-terminal fragment is released into host cell cytoplasm and is immunogenic in humans. Infect Immun 2011 Mar. 21.
23. Chen et al. The hypothetical protein CT813 is localized in the Chlamydia trachomatis inclusion membrane and is immunogenic in women urogenitally infected with C. trachomatis. Infect Immun 2006 August; 74(8):4826-40.
24. Fling et al. CD8+ T cells recognize an inclusion membrane-associated protein from the vacuolar pathogen Chlamydia trachomatis. Proc Natl Acad Sci USA 2001 Jan. 30; 98(3):1160-5.
25. Hower et al. Evidence that CT694 is a novel Chlamydia trachomatis T3 S substrate capable of functioning during invasion or early cycle development. Mol Microbiol 2009 June; 72(6):1423-37.
26. Cappello et al. Chlamydia trachomatis infection and anti-Hsp60 immunity: the two sides of the coin. PLoS Pathog 2009 August; 5(8):e1000552.
27. Domeika et al. Humoral immune response to conserved epitopes of Chlamydia trachomatis and human 60-kDa heat-shock protein in women with pelvic inflammatory disease. J Infect Dis 1998 March; 177(3):714-9.
28. Ausiello et al. 60-kDa heat shock protein of Chlamydia pneumoniae is a target of T-cell immune response. J Biol Regul Homeost Agents 2005 July-December; 19(3-4):136-40.
29. Kinnunen et al. Heat shock protein 60 specific T-cell response in chlamydial infections. Scand J Immunol 2001 July-August; 54(1-2):76-81.
30. Hjelholt et al. Tubal factor infertility is associated with antibodies against Chlamydia trachomatis heat shock protein 60 (HSP60) but not human HSP60. Hum Reprod 2011 August; 26(8):2069-76.
31. Bulut Y et al. Chlamydial heat shock protein 60 activates macrophages and endothelial cells through Toll-like receptor 4 and MD2 in a MyD88-dependent pathway. J Immunol 2002 Feb. 1; 168(3):1435-40.
32. Sharma et al. Human antibody responses to a Chlamydia-secreted protease factor. Infect Immun 2004 December; 72(12):7164-71.
33. Li et al. Characterization of fifty putative inclusion membrane proteins encoded in the Chlamydia trachomatis genome. Infect Immun 2008 June; 76(6):2746-57.
34. Gervassi et al. Human CD8+ T cells recognize the 60-kDa cysteine-rich outer membrane protein from Chlamydia trachomatis. J Immunol 2004 Dec. 1; 173(11):6905-13.

TABLE 1

Titers of antibodies against C. trachomatis and C. pneumoniae
in infertile women with or without tubal pathology

Antibodies to C. trachomatis

Antibodies to C. pneumoniae

	TFI (n = 31)	IFC (n = 23)	TFI (n = 31)	IFC (n = 23)

Quantitative	Mean	69928	3814	41503	25861
analyses	Standard	106709	8270	65848	35847
	Deviation

	Student's	p = 0.0009	p = 0.269
	t Test

Categorization of serum samples into Negative, Low and High titer groups

Qualitative	Negative	1 (3.2%)	3 (13.0%)	3 (9.7%)	4 (17.4%)
analyses	titers
	(<1:10)
	Low titers	11 (35.5%)	19 (82.6%)	11 (34.5%)	6 (26.1%)
	(1:10-
	1:10000)
	High titers	19 (61.3%)	1 (4.4%)	17 (54.8%)	13 (56.5%)
	(>1:10000)

χ²Test

p < 0.001

p = 0.634

Pairwise	High vs. Negative	p = 0.008	N/A
χ²Tests
	High vs. Low	p < 0.001
	Low vs. Negative	p = 0.556

TABLE 2

Reactivity of 30 significant C. trachomatis proteins with 31 TFI and 23 IFC patient antisera (1:800 dilution)

CT	TFI (n = 31)	IFC (n = 23)
ORF	X +/− SD	X +/− SD	t-Test	Specificity	Sensitivity	PPV	NPV

CT067	0.407 ± 0.483	0.045 ± 0.115	<0.001	87.0%	51.6%	84.2%	57.1%
CT089	0.645 ± 0.936	0.189 ± 0.407	0.020	78.3%	45.2%	73.7%	51.4%
CT110	0.679 ± 0.756	0.069 ± 0.112	<0.001	82.6%	71.0%	84.6%	67.9%
CT116	0.176 ± 0.252	0.012 ± 0.059	0.001	95.7%	32.3%	90.9%	51.2%
CT119	0.375 ± 0.501	0.082 ± 0.196	0.005	87.0%	48.4%	83.3%	55.6%
CT142	0.468 ± 0.522	0.098 ± 0.138	0.001	78.3%	54.8%	77.3%	56.3%
CT143	1.012 ± 0.818	0.166 ± 0.220	<0.001	73.9%	71.0%	78.6%	65.4%
CT147	0.789 ± 0.678	0.303 ± 0.185	0.001	34.8%	80.6%	62.5%	57.1%
CT153	0.404 ± 0.561	0.071 ± 0.110	0.003	91.3%	45.2%	87.5%	55.3%
CT322	0.366 ± 0.586	0.055 ± 0.112	0.007	95.7%	41.9%	92.9%	55.0%
CT376	0.453 ± 0.616	0.072 ± 0.097	0.002	95.7%	41.9%	92.9%	55.0%
CT381	0.330 ± 0.346	0.059 ± 0.074	<0.001	95.7%	51.6%	94.1%	59.5%
CT414	0.327 ± 0.469	0.061 ± 0.082	0.004	95.7%	51.6%	94.1%	59.5%
CT442	0.486 ± 0.622	0.055 ± 0.070	0.001	91.3%	48.4%	88.2%	56.8%
CT443	1.145 ± 1.020	0.110 ± 0.173	<0.001	87.0%	71.0%	88.0%	69.0%
CT456	0.803 ± 0.879	0.241 ± 0.558	0.006	73.9%	64.5%	76.9%	60.7%
CT529	0.854 ± 0.644	0.444 ± 0.310	0.003	13.0%	87.1%	57.4%	42.9%
CT557	0.358 ± 0.638	0.028 ± 0.057	0.007	100%	29.0%	100%	51.1%
CT603	0.579 ± 0.654	0.141 ± 0.161	0.001	78.3%	61.3%	79.2%	60.0%
CT681	0.363 ± 0.386	0.060 ± 0.078	<0.001	87.0%	51.6%	84.2%	57.1%
CT694	0.698 ± 0.848	0.150 ± 0.293	0.002	78.3%	54.8%	77.3%	56.3%
CT795	0.647 ± 0.771	0.034 ± 0.101	<0.001	95.7%	61.3%	95.0%	64.7%
CT798	0.622 ± 0.827	0.038 ± 0.089	<0.001	91.3%	51.6%	88.9%	58.3%
CT806	0.673 ± 0.772	0.104 ± 0.212	<0.001	82.6%	54.8%	81.0%	57.6%
CT812	0.555 ± 0.667	0.061 ± 0.087	<0.001	91.3%	54.8%	89.5%	60.0%
CT813	0.673 ± 0.689	0.095 ± 0.165	<0.001	78.3%	67.7%	80.8%	64.3%
CT823	0.649 ± 0.709	0.071 ± 0.074	<0.001	91.3%	71.0%	91.7%	70.0%
CT858	1.947 ± 1.276	0.338 ± 0.666	<0.001	78.3%	74.2%	82.1%	69.2%
CT866	0.574 ± 0.738	0.062 ± 0.079	0.001	91.3%	48.4%	88.2%	56.8%
pCT03	1.761 ± 1.366	0.166 ± 0.573	<0.001	82.6%	74.2%	85.2%	70.4%

TABLE 3

Titers of antibodies against C. trachomatis and C. pneumoniae
in women with tubal factor infertility (TFI), normal fertility
(FC) or acute infection (STI)

Antibodies to C. trachomatis

Antibodies to C. pneumoniae

	TFI	FC	STI	TFI	FC	STI
Groups	(n = 24)	(n = 25)	(n = 24)	(n = 24)	(n = 25)	(n = 24)

Mean	35483.33	23760.00	65500.00	23000.00	17136.00	7875.00
Standard	39950.49	23974.43	70078.87	23083.97	19709.45	6295.70

deviation
ANOVA	PP = 0.0108	PP = 0.0166

	TFI vs	FC vs.	TFI vs.	TFI vs	FC vs	TFI vs.
	FC	STI	STI	FC	STI	STI

Student's	P = 0.2229	P = 0.0099	P = 0.0748	P = 0.3431	PP = 0.0331	P = 0.0033
t-Test

TABLE 4

Antigens preferentially recognized by woman with Tubal Factor
Infertility but not normal fertility

Groups

TFI (N = 24)

FC (N = 25)

P value

ORFs	Freq.	Mean ± SD	Freq.	Mean ± SD	Fisher's	t-Test

CT110
	38%	0.308 ± 0.512	0%	0.018 ± 0.054	<0.001	0.011
CT376	25%	0.137 ± 0.186	0%	0.005 ± 0.037	0.010	0.001
CT111	17%	0.129 ± 0.238	0%	0.043 ± 0.062	0.050	0.098
CT557	17%	0.142 ± 0.289	0%	0.008 ± 0.047	0.050	0.035
CT579	17%	0.077 ± 0.163	0%	0.004 ± 0.040	0.050	0.025
CT443	88%	0.717 ± 0.576	60%	0.306 ± 0.258	0.030	0.003
CT798	58%	0.374 ± 0.403	28%	0.182 ± 0.328	0.031	0.074
CT603	42%	0.187 ± 0.203	16%	0.071 ± 0.140	0.047	0.025
CT381	21%	0.102 ± 0.149	4%	0.027 ± 0.072	0.086	0.031
CT823	17%	0.122 ± 0.158	4%	0.034 ± 0.109	0.162	0.029

TABLE 5

21 antigens most significantly recognized by women with acute C. trachomatis infection
(antiserum dilution 1:1.000)

Groups

STI (n = 24)

TFI (n = 24)

FC (n = 25)

STI vs. TFI

STI vs. FC

TFI vs. FC

ORFs

Freq

Mean ± SD

Freq

Mean ± SD

Freq

Mean ± SD

t-Test

Fisher's

t-Test

Fisher's

t-Test

Fisher's

CT147	88%	0.937 ± 0.776	21%	0.161 ± 0.191	16%	0.117 ± 0.213	<0.001	<0.001	<0.001	<0.001	0.455	0.661
CT442	83%	0.841 ± 0.701	17%	0.151 ± 0.303	20%	0.113 ± 0.229	<0.001	<0.001	<0.001	<0.001	0.623	0.763
CT529	100%	1.040 ± 0.620	42%	0.293 ± 0.310	32%	0.227 ± 0.341	<0.001	<0.001	<0.001	<0.001	0.483	0.482
CT119	75%	0.603 ± 0.681	13%	0.124 ± 0.212	4%	0.040 ± 0.072	0.002	<0.001	<0.001	<0.001	0.077	0.277
CT089	92%	1.364 ± 0.995	33%	0.274 ± 0.456	28%	0.208 ± 0.388	<0.001	<0.001	<0.001	<0.001	0.586	0.685
CT695	92%	0.825 ± 0.731	33%	0.274 ± 0.342	24%	0.144 ± 0.149	0.002	<0.001	<0.001	<0.001	0.098	0.469
CT806	88%	0.739 ± 0.643	29%	0.152 ± 0.231	20%	0.102 ± 0.152	<0.001	<0.001	<0.001	<0.001	0.387	0.455
CT875	75%	0.956 ± 0.727	17%	0.145 ± 0.269	16%	0.098 ± 0.170	<0.001	<0.001	<0.001	<0.001	0.472	0.949
CT841	96%	1.093 ± 0.625	42%	0.282 ± 0.468	48%	0.232 ± 0.255	<0.001	<0.001	<0.001	<0.001	0.643	0.655
CT694	92%	1.235 ± 0.763	38%	0.261 ± 0.375	32%	0.311 ± 0.505	<0.001	<0.001	<0.001	<0.001	0.695	0.685
CT480	54%	0.447 ± 0.529	4%	0.046 ± 0.143	8%	0.027 ± 0.101	<0.001	<0.001	<0.001	<0.001	0.594	0.575
CT812	63%	0.654 ± 0.731	13%	0.137 ± 0.286	4%	0.028 ± 0.075	0.002	<0.001	<0.001	<0.001	0.083	0.277
CT228	50%	0.377 ± 0.675	4%	0.037 ± 0.063	0%	0.009 ± 0.041	0.018	<0.001	0.013	<0.001	0.078	0.302
CT153	83%	0.765 ± 0.785	33%	0.238 ± 0.312	16%	0.112 ± 0.134	0.004	<0.001	<0.001	<0.001	0.079	0.158
CT381	71%	0.537 ± 0.579	21%	0.102 ± 0.148	4%	0.026 ± 0.071	<0.001	<0.001	<0.001	<0.001	0.031	0.072
CT118	46%	0.298 ± 0.494	4%	0.041 ± 0.077	0%	0.018 ± 0.046	0.016	<0.001	0.011	<0.001	0.216	0.302
CT149	67%	0.594 ± 0.941	21%	0.136 ± 0.285	12%	0.067 ± 0.192	0.027	0.001	0.012	<0.001	0.331	0.402
CT142	92%	1.173 ± 0.686	50%	0.265 ± 0.310	28%	0.204 ± 0.337	<0.001	0.001	<0.001	<0.001	0.512	0.114
CT619	33%	0.184 ± 0.428	0%	0.005 ± 0.035	4%	0.003 ± 0.054	0.047	0.002	0.050	0.008	0.866	0.322
CT702	33%	0.279 ± 0.649	0%	0.017 ± 0.038	4%	0.015 ± 0.067	0.030	0.002	0.037	0.008	0.875	0.322
CT143	100%	1.324 ± 0.681	67%	0.355 ± 0.317	40%	0.202 ± 0.219	<0.001	0.002	<0.001	<0.001	0.058	0.060

TABLE 6

5-mer CT443 (OmcB or CRP60, 553 amino acids),
ACCESSION#: NP219955.1, GI: 15605169 (SEQ ID NOS:
1-549)

1.	MRIGD
2.	RIGDP
3.	IGDPM
4.	GDPMN
5.	DPMNK
6.	PMNKL
7.	MNKLI
8.	NKLIR
9.	KLIRR
10.	LIRRA
11.	IRRAV
12.	RRAVT
13.	RAVTI
14.	AVTIF
15.	VTIFA
16.	TIFAV
17.	IFAVT
18.	FAVTS
19.	AVTSV
20.	VTSVA
21.	TSVAS
22.	SVASL
23.	VASLF
24.	ASLFA
25.	SLFAS
26.	LFASG
27.	FASGV
28.	ASGVL
29.	SGVLE
30.	GVLET
31.	VLETS
32.	LETSM
33.	ETSMA
34.	TSMAE
35.	SMAES
36.	MAESL
37.	AESLS
38.	ESLST
39.	SLSTN
40.	YATVG
41.	STNVI
42.	TNVIS
43.	NVISL
44.	VISLA
45.	ISLAD
46.	SLADT
47.	LADTK
48.	ADTKA
49.	DTKAK
50.	TKAKD
51.	KAKDN
52.	AKDNT
53.	KDNTS
54.	DNTSH
55.	NTSHK
56.	TSHKS
57.	SHKSK
58.	HKSKK
59.	KSKKA
60.	SKKAR
61.	KKARK
62.	KARKN
63.	ARKNH
64.	RKNHS
65.	KNHSK
66.	NHSKE
67.	HSKET
68.	SKETP
69.	KETPV
70.	ETPVD
71.	TPVDR
72.	PVDRK
73.	VDRKE
74.	DRKEV
75.	RKEVA
76.	KEVAP
77.	EVAPV
78.	VAPVH
79.	APVHE
80.	PVHES
81.	TTPTA
82.	HESKA
83.	ESKAT
84.	SKATG
85.	KATGP
86.	ATGPK
87.	TGPKQ
88.	GPKQD
89.	PKQDS
90.	KQDSC
91.	QDSCF
92.	DSCFG
93.	SCFGR
94.	CFGRM
95.	FGRMY
96.	GRMYT
97.	RMYTV
98.	MYTVK
99.	YTVKV
100.	TVKVN
101.	VKVND
102.	KVNDD
103.	VNDDR
104.	NDDRN
105.	DDRNV
106.	DRNVE
107.	RNVEI
108.	NVEIT
109.	VEITQ
110.	EITQA
111.	ITQAV
112.	TQAVP
113.	QAVPE
114.	AVPEY
115.	VPEYA
116.	PEYAT
117.	EYATV
118.	YATVG
119.	ATVGS
120.	TVGSP
121.	VGSPY
122.	GSPYP
123.	SPYPI
124.	PYPIE
125.	YPIEI
126.	PIEIT
127.	IEITA
128.	EITAT
129.	ITATG
130.	TATGK
131.	ATGKR
132.	TGKRD
133.	GKRDC
134.	KRDCV
135.	RDCVD
136.	DCVDV
137.	CVDVI
138.	VDVII
139.	DVIIT
140.	VIITQ
141.	IITQQ
142.	ITQQL
143.	TQQLP
144.	QQLPC
145.	QLPCE
146.	LPCEA
147.	PCEAE
148.	CEAEF
149.	EAEFV
150.	AEFVR
151.	EFVRS
152.	FVRSD
153.	VRSDP
154.	RSDPA
155.	SDPAT
156.	DPATT
157.	PATTP
158.	ATTPT
159.	TTPTA
160.	TPTAD
161.	PTADG
162.	TADGK
163.	ADGKL
164.	DGKLV
165.	GKLVW
166.	KLVWK
167.	LVWKI
168.	VWKID
169.	WKIDR
170.	KIDRL
171.	IDRLG
172.	DRLGQ
173.	RLGQG
174.	LGQGE
175.	GQGEK
176.	QGEKS
177.	GEKSK
178.	EKSKI
179.	KSKIT
180.	SKITV
181.	KITVW
182.	ITVWV
183.	TVWVK
184.	VWVKP
185.	WVKPL
186.	VKPLK
187.	KPLKE
188.	PLKEG
189.	LKEGC
190.	KEGCC
191.	EGCCF
192.	GCCFT
193.	CCFTA
194.	CFTAA
195.	FTAAT
196.	TAATV
197.	AATVC
198.	ATVCA
199.	TVCAC
200.	VCACP
201.	CACPE
202.	ACPEI
203.	CPEIR
204.	PEIRS
205.	EIRSV
206.	IRSVT
207.	RSVTK
208.	SVTKC
209.	VTKCG
210.	TKCGQ
211.	KCGQP
212.	CGQPA
213.	GQPAI
214.	QPAIC
215.	PAICV
216.	AICVK
217.	ICVKQ
218.	CVKQE
219.	VKQEG
220.	KQEGP
221.	QEGPE
222.	EGPEN
223.	GPENA
224.	PENAC
225.	ENACL
226.	NACLR
227.	ACLRC
228.	CLRCP
229.	LRCPV
230.	RCPVV
231.	CPVVY
232.	PVVYK
233.	VVYKI
234.	VYKIN
235.	YKINI
236.	KINIV
237.	INIVN
238.	NIVNQ
239.	IVNQG
240.	VNQGT
241.	NQGTA
242.	QGTAT
243.	GTATA
244.	TATAR
245.	ATARN
246.	TARNV
247.	ARNVV
248.	RNVVV
249.	NVVVE
250.	VVVEN
251.	VVENP
252.	VENPV
253.	ENPVP
254.	NPVPD
255.	PVPDG
256.	VPDGY
257.	PDGYA
258.	DGYAH
259.	GYAHS
260.	YAHSS
261.	AHSSG
262.	HSSGQ
263.	SSGQR
264.	SGQRV
265.	GQRVL
266.	QRVLT
267.	RVLTF
268.	VLTFT
269.	LTFTL
270.	TFTLG
271.	FTLGD
272.	TLGDM
273.	LGDMQ
274.	GDMQP
275.	DMQPG
276.	MQPGE
277.	QPGEH
278.	PGEHR
279.	GEHRT
280.	EHRTI
281.	HRTIT
282.	RTITV
283.	TITVE
284.	ITVEF
285.	TVEFC
286.	VEFCP
287.	EFCPL
288.	FCPLK
289.	CPLKR
290.	PLKRG
291.	LKRGR
292.	KRGRA
293.	RGRAT
294.	GRATN
295.	RATNI
296.	ATNIA
297.	TNIAT
298.	NIATV
299.	IATVS
300.	ATVSY
301.	TVSYC
302.	VSYCG
303.	SYCGG
304.	YCGGH
305.	CGGHK
306.	GGHKN
307.	GHKNT
308.	HKNTA
309.	KNTAS
310.	NTASV
311.	TASVT
312.	ASVTT
313.	SVTTV
314.	VTTVI
315.	TTVIN
316.	TVINE
317.	VINEP
318.	INEPC
319.	NEPCV
320.	EPCVQ
321.	PCVQV
322.	CVQVS
323.	VQVSI
324.	QVSIA
325.	VSIAG
326.	SIAGA
327.	IAGAD
328.	AGADW
329.	GADWS
330.	ADWSY
331.	DWSYV
332.	WSYVC
333.	SYVCK
334.	YVCKP
335.	VCKPV
336.	CKPVE
337.	KPVEY
338.	PVEYV
339.	VEYVI
340.	EYVIS
341.	YVISV
342.	VISVS
343.	ISVSN
344.	SVSNP
345.	VSNPG
346.	SNPGD
347.	NPGDL
348.	PGDLV
349.	GDLVL
350.	DLVLR
351.	LVLRD
352.	VLRDV
353.	LRDVV
354.	RDVVV
355.	DVVVE
356.	VVVED
357.	VVEDT
358.	VEDTL
359.	EDTLS
360.	DTLSP
361.	TLSPG
362.	LSPGV
363.	SPGVT
364.	PGVTV
365.	GVTVL
366.	VTVLE
367.	TVLEA
368.	VLEAA
369.	LEAAG
370.	EAAGA
371.	AAGAQ
372.	AGAQI
373.	GAQIS
374.	AQISC
375.	QISCN
376.	ISCNK
377.	SCNKV
378.	CNKVV
379.	NKVVW
380.	KVVWT
381.	VVWTV
382.	VWTVK
383.	WTVKE
384.	TVKEL
385.	VKELN
386.	KELNP
387.	ELNPG
388.	LNPGE
389.	NPGES
390.	PGESL
391.	GESLQ
392.	ESLQY
393.	SLQYK
394.	LQYKV
395.	QYKVL
396.	YKVLV
397.	KVLVR
398.	VLVRA
399.	LVRAQ
400.	VRAQT
401.	RAQTP
402.	AQTPG
403.	QTPGQ
404.	TPGQF
405.	PGQFT
406.	GQFTN
407.	QFTNN
408.	FTNNV
409.	TNNVV
410.	NNVVV
411.	NVVVK
412.	VVVKS
413.	VVKSC
414.	VKSCS
415.	KSCSD
416.	SCSDC
417.	CSDCG
418.	SDCGT
419.	DCGTC
420.	CGTCT
421.	GTCTS
422.	TCTSC
423.	CTSCA
424.	TSCAE
425.	SCAEA
426.	CAEAT
427.	AEATT
428.	EATTY
429.	ATTYW
430.	TTYWK
431.	TYWKG
432.	YWKGV
433.	WKGVA
434.	KGVAA
435.	GVAAT
436.	VAATH
437.	AATHM
438.	ATHMC
439.	THMCV
440.	HMCVV
441.	MCVVD
442.	CVVDT
443.	VVDTC
444.	VDTCD
445.	DTCDP
446.	TCDPV
447.	CDPVC
448.	DPVCV
449.	PVCVG
450.	VCVGE
451.	CVGEN
452.	VGENT
453.	GENTV
454.	ENTVY
455.	NTVYR
456.	TVYRI
457.	VYRIC
458.	YRICV
459.	RICVT
460.	ICVTN
461.	CVTNR
462.	VTNRG
463.	TNRGS
464.	NRGSA
465.	RGSAE
466.	GSAED
467.	SAEDT
468.	AEDTN
469.	EDTNV
470.	DTNVS
471.	TNVSL
472.	NVSLM
473.	VSLML
474.	SLMLK
475.	LMLKF
476.	MLKFS
477.	LKFSK
478.	KFSKE
479.	FSKEL
480.	SKELQ
481.	KELQP
482.	ELQPV
483.	LQPVS
484.	QPVSF
485.	PVSFS
486.	VSFSG
487.	SFSGP
488.	FSGPT
489.	SGPTK
490.	GPTKG
491.	PTKGT
492.	TKGTI
493.	KGTIT
494.	GTITG
495.	TITGN
496.	ITGNT
497.	TGNTV
498.	GNTVV
499.	NTVVF
500.	TVVFD
501.	VVFDS
502.	VFDSL
503.	FDSLP
504.	DSLPR
505.	SLPRL
506.	LPRLG
507.	PRLGS
508.	RLGSK
509.	LGSKE
510.	GSKET
511.	SKETV
512.	KETVE
513.	ETVEF
514.	TVEFS
515.	VEFSV
516.	EFSVT
517.	FSVTL
518.	SVTLK
519.	VTLKA
520.	TLKAV
521.	LKAVS
522.	KAVSA
523.	AVSAG
524.	VSAGD
525.	SAGDA
526.	AGDAR
527.	GDARG
528.	DARGE
529.	ARGEA
530.	RGEAI
531.	GEAIL
532.	EAILS
533.	AILSS
534.	ILSSD
535.	LSSDT
536.	SSDTL
537.	SDTLT
538.	DTLTV
539.	TLTVP
540.	LTVPV
541.	TVPVS
542.	VPVSD
543.	PVSDT
544.	VSDTE
545.	SDTEN
546.	DTENT
547.	TENTH
548.	ENTHI
549.	NTHIY

5-mer peptides of CT381 (ArtJ, 257 amino acids),

accession#: NP 219890.1, GI: 15605105 (SEQ ID

NOS: 550-802)

550.	MCIKR
551.	CIKRK
552.	IKRKK
553.	KRKKT
554.	RKKTW
555.	KKTWI
556.	KTWIA
557.	TWIAF
558.	WIAFL
559.	IAFLA
560.	AFLAV
561.	FLAVV
562.	LAVVC
563.	AVVCS
564.	VVCSF
565.	VCSFC
566.	CSFCL
567.	SFCLT
568.	FCLTG
569.	CLTGC
570.	LTGCL
571.	TGCLK
572.	GCLKE
573.	CLKEG
574.	LKEGG
575.	KEGGD
576.	EGGDS
577.	GGDSN
578.	GDSNS
579.	DSNSE
580.	SNSEK
581.	NSEKF
582.	SEKFI
583.	EKFIV
584.	KFIVG
585.	FIVGT
586.	IVGTN
587.	VGTNA
588.	GTNAT
589.	TNATY
590.	NATYP
591.	ATYPP
592.	TYPPF
593.	YPPFE
594.	PPFEF
595.	PFEFV
596.	FEFVD
597.	EFVDK
598.	FVDKR
599.	VDKRG
600.	DKRGE
601.	KRGEV
602.	RGEVV
603.	GEVVG
604.	EVVGF
605.	VVGFD
606.	VGFDI
607.	GFDID
608.	FDIDL
609.	DIDLA
610.	IDLAR
611.	DLARE
612.	LAREI
613.	AREIS
614.	REISN
615.	EISNK
616.	ISNKL
617.	SNKLG
618.	NKLGK
619.	KLGKT
620.	LGKTL
621.	GKTLD
622.	KTLDV
623.	TLDVR
624.	LDVRE
625.	DVREF
626.	VREFS
627.	REFSF
628.	EFSFD
629.	FSFDA
630.	SFDAL
631.	FDALI
632.	DALIL
633.	ALILN
634.	LILNL
635.	ILNLK
636.	LNLKQ
637.	NLKQH
638.	LKQHR
639.	KQHRI
640.	QHRID
641.	HRIDA
642.	RIDAV
643.	IDAVI
644.	DAVIT
645.	AVITG
646.	VITGM
647.	ITGMS
648.	TGMSI
649.	GMSIT
650.	MSITP
651.	SITPS
652.	ITPSR
653.	TPSRL
654.	PSRLK
655.	SRLKE
656.	RLKEI
657.	LKEIL
658.	KEILM
659.	EILMI
660.	ILMIP
661.	LMIPY
662.	MIPYY
663.	IPYYG
664.	PYYGE
665.	YYGEE
666.	YGEEI
667.	GEEIK
668.	EEIKH
669.	EIKHL
670.	IKHLV
671.	KHLVL
672.	HLVLV
673.	LVLVF
674.	VLVFK
675.	LVFKG
676.	VFKGE
677.	FKGEN
678.	KGENK
679.	GENKH
680.	ENKHP
681.	NKHPL
682.	KHPLP
683.	HPLPL
684.	PLPLT
685.	LPLTQ
686.	PLTQY
687.	LTQYR
688.	TQYRS
689.	QYRSV
690.	YRSVA
691.	RSVAV
692.	SVAVQ
693.	VAVQT
694.	AVQTG
695.	VQTGT
696.	QTGTY
697.	TGTYQ
698.	GTYQE
699.	TYQEA
700.	YQEAY
701.	QEAYL
702.	EAYLQ
703.	AYLQS
704.	YLQSL
705.	LQSLS
706.	QSLSE
707.	SLSEV
708.	LSEVH
709.	SEVHI
710.	EVHIR
711.	VHIRS
712.	HIRSF
713.	IRSFD
714.	RSFDS
715.	SFDST
716.	FDSTL
717.	DSTLE
718.	STLEV
719.	TLEVL
720.	LEVLM
721.	EVLME
722.	VLMEV
723.	LMEVM
724.	MEVMH
725.	EVMHG
726.	VMHGK
727.	MHGKS
728.	HGKSP
729.	GKSPV
730.	KSPVA
731.	SPVAV
732.	PVAVL
733.	VAVLE
734.	AVLEP
735.	VLEPS
736.	LEPSI
737.	EPSIA
738.	PSIAQ
739.	SIAQV
740.	IAQVV
741.	AQVVL
742.	QVVLK
743.	VVLKD
744.	VLKDF
745.	LKDFP
746.	KDFPA
747.	DFPAL
748.	FPALS
749.	PALST
750.	ALSTA
751.	LSTAT
752.	STATI
753.	TATID
754.	ATIDL
755.	TIDLP
756.	IDLPE
757.	DLPED
758.	LPEDQ
759.	PEDQW
760.	EDQWV
761.	DQWVL
762.	QWVLG
763.	WVLGY
764.	VLGYG
765.	LGYGI
766.	GYGIG
767.	YGIGV
768.	GIGVA
769.	IGVAS
770.	GVASD
771.	VASDR
772.	ASDRP
773.	SDRPA
774.	DRPAL
775.	RPALA
776.	PALAL
777.	ALALK
778.	LALKI
779.	ALKIE
780.	LKIEA
781.	KIEAA
782.	IEAAV
783.	EAAVQ
784.	AAVQE
785.	AVQEI
786.	VQEIR
787.	QEIRK
788.	EIRKE
789.	IRKEG
790.	RKEGV
791.	KEGVL
792.	EGVLA
793.	GVLAE
794.	VLAEL
795.	LAELE
796.	AELEQ
797.	ELEQK
798.	LEQKW
799.	EQKWG
800.	QKWGL
801.	KWGLN
802.	WGLNN

5-mer peptides of CT875 (hypothetical protein, 591

amino acids), ACCESSION#: NP 219502.1, GI:

15604718 (SEQ ID NOS: 803-1389)

803.	MSIRG
804.	SIRGV
805.	IRGVG
806.	RGVGG
807.	GVGGN
808.	VGGNG
809.	GNGNS
810.	NGNSR
811.	GNSRI
812.	NSRIP
813.	SRIPS
814.	RIPSH
815.	IPSHN
816.	PSHNG
817.	SHNGD
818.	HNGDG
819.	NGDGS
820.	GDGSN
821.	DGSNR
822.	GSNRR
823.	SNRRS
824.	NRRSQ
825.	RRSQN
826.	RSQNT
827.	SQNTK
828.	QNTKG
829.	NTKGN
830.	TKGNN
831.	KGNNK
832.	GNNKV
833.	NNKVE
834.	NKVED
835.	KVEDR
836.	VEDRV
837.	VEDRV
838.	EDRVC
839.	DRVCS
840.	RVCSL
841.	VCSLY
842.	CSLYS
843.	SLYSS
844.	LYSSR
845.	YSSRS
846.	SSRSN
847.	SRSNE
848.	RSNEN
849.	SNENR
850.	NENRE
851.	ENRES
852.	NRESP
853.	RESPY
854.	ESPYA
855.	SPYAV
856.	PYAVV
857.	YAVVD
858.	AVVDV
859.	VVDVS
860.	VDVSS
861.	DVSSM
862.	VSSMI
863.	SSMIE
864.	SMIES
865.	MIEST
866.	IESTP
867.	ESTPT
868.	STPTS
869.	TPTSG
870.	PTSGE
871.	TSGET
872.	SGETT
873.	GETTR
874.	ETTRA
875.	TTRAS
876.	TRASR
877.	RASRG
878.	ASRGV
879.	SRGVF
880.	RGVFS
881.	GVFSR
882.	VFSRF
883.	FSRFQ
884.	SRFQR
885.	RFQRG
886.	FQRGL
887.	QRGLV
888.	RGLVR
889.	GLVRV
890.	LVRVA
891.	VRVAD
892.	RVADK
893.	VADKV
894.	ADKVR
895.	DKVRR
896.	KVRRA
897.	VRRAV
898.	RRAVQ
899.	RAVQC
900.	AVQCA
901.	VQCAW
902.	QCAWS
903.	CAWSS
904.	AWSSV
905.	WSSVS
906.	SSVST
907.	SVSTR
908.	VSTRR
909.	STRRS
910.	TRRSS
911.	RRSSA
912.	RSSAT
913.	SSATR
914.	SATRA
915.	ATRAA
916.	TRAAE
917.	RAAES
918.	AAESG
919.	AESGS
920.	ESGSS
921.	SGSSS
922.	GSSSR
923.	SSSRT
924.	SSRTA
925.	SRTAR
926.	RTARG
927.	TARGA
928.	ARGAS
929.	RGASS
930.	GASSG
931.	ASSGY
932.	SSGYR
933.	SGYRE
934.	GYREY
935.	YREYS
936.	REYSP
937.	EYSPS
938.	YSPSA
939.	SPSAA
940.	PSAAR
941.	SAARG
942.	AARGL
943.	ARGLR
944.	RGLRL
945.	GLRLM
946.	LRLMF
947.	RLMFT
948.	LMFTD
949.	MFTDF
950.	FTDFW
951.	TDFWR
952.	DFWRT
953.	FWRTR
954.	WRTRV
955.	RTRVL
956.	TRVLR
957.	RVLRQ
958.	VLRQT
959.	LRQTS
960.	RQTSP
961.	QTSPM
962.	TSPMA
963.	SPMAG
964.	PMAGV
965.	MAGVF
966.	AGVFG
967.	GVFGN
968.	VFGNL
969.	FGNLD
970.	GNLDV
971.	NLDVN
972.	LDVNE
973.	DVNEA
974.	VNEAR
975.	NEARL
976.	EARLM
977.	ARLMA
978.	RLMAA
979.	LMAAY
980.	MAAYT
981.	AAYTS
982.	AYTSE
983.	YTSEC
984.	TSECA
985.	SECAD
986.	ECADH
987.	CADHL
988.	ADHLE
989.	DHLEA
990.	HLEAN
991.	LEANK
992.	EANKL
993.	ANKLA
994.	NKLAG
995.	KLAGP
996.	LAGPD
997.	AGPDG
998.	GPDGV
999.	PDGVA
1000.	DGVAA
1001.	GVAAA
1002.	VAAAR
1003.	AAARE
1004.	AAREI
1005.	AREIA
1006.	REIAK
1007.	EIAKR
1008.	IAKRW
1009.	AKRWE
1010.	KRWEQ
1011.	RWEQR
1012.	WEQRV
1013.	EQRVR
1014.	QRVRD
1015.	RVRDL
1016.	VRDLQ
1017.	RDLQD
1018.	DLQDK
1019.	LQDKG
1020.	QDKGA
1021.	DKGAA
1022.	KGAAR
1023.	GAARK
1024.	AARKL
1025.	ARKLL
1026.	RKLLN
1027.	KLLND
1028.	LLNDP
1029.	LNDPL
1030.	NDPLG
1031.	DPLGR
1032.	PLGRR
1033.	LGRRT
1034.	GRRTP
1035.	RRTPN
1036.	RTPNY
1037.	TPNYQ
1038.	PNYQS
1039.	NYQSK
1040.	YQSKN
1041.	QSKNP
1042.	SKNPG
1043.	KNPGE
1044.	NPGEY
1045.	PGEYT
1046.	GEYTV
1047.	EYTVG
1048.	YTVGN
1049.	TVGNS
1050.	VGNSM
1051.	GNSMF
1052.	NSMFY
1053.	SMFYD
1054.	MFYDG
1055.	FYDGP
1056.	YDGPQ
1057.	DGPQV
1058.	GPQVA
1059.	PQVAN
1060.	QVANL
1061.	VANLQ
1062.	ANLQN
1063.	NLQNV
1064.	LQNVD
1065.	QNVDT
1066.	NVDTG
1067.	VDTGF
1068.	DTGFW
1069.	TGFWL
1070.	GFWLD
1071.	FWLDM
1072.	WLDMS
1073.	LDMSN
1074.	DMSNL
1075.	MSNLS
1076.	SNLSD
1077.	NLSDV
1078.	LSDVV
1079.	SDVVL
1080.	DVVLS
1081.	VVLSR
1082.	VLSRE
1083.	LSREI
1084.	SREIQ
1085.	REIQT
1086.	EIQTG
1087.	IQTGL
1088.	QTGLR
1089.	TGLRA
1090.	GLRAR
1091.	LRARA
1092.	RARAT
1093.	ARATL
1094.	RATLE
1095.	ATLEE
1096.	TLEES
1097.	LEESM
1098.	EESMP
1099.	ESMPM
1100.	SMPML
1101.	MPMLE
1102.	PMLEN
1103.	MLENL
1104.	LENLE
1105.	ENLEE
1106.	NLEER
1107.	LEERF
1108.	EERFR
1109.	ERFRR
1110.	RFRRL
1111.	FRRLQ
1112.	RRLQE
1113.	RLQET
1114.	LQETC
1115.	QETCD
1116.	ETCDA
1117.	TCDAA
1118.	CDAAR
1119.	DAART
1120.	AARTE
1121.	ARTEI
1122.	RTEIE
1123.	TEIEE
1124.	EIEES
1125.	IEESG
1126.	EESGW
1127.	ESGWT
1128.	SGWTR
1129.	GWTRE
1130.	WTRES
1131.	TRESA
1132.	RESAS
1133.	ESASR
1134.	SASRM
1135.	ASRME
1136.	SRMEG
1137.	RMEGD
1138.	MEGDE
1139.	EGDEA
1140.	GDEAQ
1141.	DEAQG
1142.	EAQGP
1143.	AQGPS
1144.	QGPSR
1145.	GPSRA
1146.	PSRAQ
1147.	SRAQQ
1148.	RAQQA
1149.	AQQAF
1150.	QQAFQ
1151.	QAFQS
1152.	AFQSF
1153.	FQSFV
1154.	QSFVN
1155.	SFVNE
1156.	FVNEC
1157.	VNECN
1158.	NECNS
1159.	ECNSI
1160.	CNSIE
1161.	NSIEF
1162.	SIEFS
1163.	IEFSF
1164.	EFSFG
1165.	FSFGS
1166.	SFGSF
1167.	FGSFG
1168.	GSFGE
1169.	SFGEH
1170.	FGEHV
1171.	GEHVR
1172.	EHVRV
1173.	HVRVL
1174.	VRVLC
1175.	RVLCA
1176.	VLCAR
1177.	LCARV
1178.	CARVS
1179.	ARVSR
1180.	RVSRG
1181.	VSRGL
1182.	SRGLA
1183.	RGLAA
1184.	GLAAA
1185.	LAAAG
1186.	AAAGE
1187.	AAGEA
1188.	AGEAI
1189.	GEAIR
1190.	EAIRR
1191.	AIRRC
1192.	IRRCF
1193.	RRCFS
1194.	RCFSC
1195.	CFSCC
1196.	FSCCK
1197.	SCCKG
1198.	CCKGS
1199.	CKGST
1200.	KGSTH
1201.	GSTHR
1202.	STHRY
1203.	THRYA
1204.	HRYAP
1205.	RYAPR
1206.	YAPRD
1207.	APRDD
1208.	PRDDL
1209.	RDDLS
1210.	DDLSP
1211.	DLSPE
1212.	LSPEG
1213.	SPEGA
1214.	PEGAS
1215.	EGASL
1216.	GASLA
1217.	ASLAE
1218.	SLAET
1219.	LAETL
1220.	AETLA
1221.	ETLAR
1222.	TLARF
1223.	LARFA
1224.	ARFAD
1225.	RFADD
1226.	FADDM
1227.	ADDMG
1228.	DDMGI
1229.	DMGIE
1230.	MGIER
1231.	GIERG
1232.	IERGA
1233.	ERGAD
1234.	RGADG
1235.	GADGT
1236.	ADGTY
1237.	DGTYD
1238.	GTYDI
1239.	TYDIP
1240.	YDIPL
1241.	DIPLV
1242.	IPLVD
1243.	PLVDD
1244.	LVDDW
1245.	VDDWR
1246.	DDWRR
1247.	DWRRG
1248.	WRRGV
1249.	RRGVP
1250.	RGVPS
1251.	GVPSI
1252.	VPSIE
1253.	PSIEG
1254.	SIEGE
1255.	IEGEG
1256.	EGEGS
1257.	GEGSD
1258.	EGSDS
1259.	GSDSI
1260.	SDSIY
1261.	DSIYE
1262.	SIYEI
1263.	IYEIM
1264.	YEIMM
1265.	EIMMP
1266.	IMMPI
1267.	MMPIY
1268.	MPIYE
1269.	PIYEV
1270.	IYEVM
1271.	YEVMD
1272.	EVMDM
1273.	VMDMD
1274.	MDMDL
1275.	DMDLE
1276.	MDLET
1277.	DLETR
1278.	LETRR
1279.	ETRRS
1280.	TRRSF
1281.	RRSFA
1282.	RSFAV
1283.	SFAVQ
1284.	FAVQQ
1285.	AVQQG
1286.	VQQGH
1287.	QQGHY
1288.	QGHYQ
1289.	GHYQD
1290.	HYQDP
1291.	YQDPR
1292.	QDPRA
1293.	DPRAS
1294.	PRASD
1295.	RASDY
1296.	ASDYD
1297.	SDYDL
1298.	DYDLP
1299.	YDLPR
1300.	DLPRA
1301.	LPRAS
1302.	PRASD
1303.	RASDY
1304.	ASDYD
1305.	SDYDL
1306.	DYDLP
1307.	YDLPR
1308.	DLPRS
1309.	LPRSP
1310.	PRSPY
1311.	RSPYP
1312.	SPYPT
1313.	PYPTP
1314.	YPTPP
1315.	PTPPL
1316.	TPPLP
1317.	PPLPP
1318.	PLPPR
1319.	LPPRY
1320.	PPRYQ
1321.	PRYQL
1322.	RYQLQ
1323.	YQLQN
1324.	QLQNM
1325.	LQNMD
1326.	QNMDV
1327.	NMDVE
1328.	MDVEA
1329.	DVEAG
1330.	VEAGF
1331.	EAGFR
1332.	AGFRE
1333.	GFREA
1334.	FREAV
1335.	REAVY
1336.	EAVYA
1337.	AVYAS
1338.	VYASF
1339.	YASFV
1340.	ASFVA
1341.	SFVAG
1342.	FVAGM
1343.	VAGMY
1344.	AGMYN
1345.	GMYNY
1346.	MYNYV
1347.	YNYVV
1348.	NYVVT
1349.	YVVTQ
1350.	VVTQP
1351.	VTQPQ
1352.	TQPQE
1353.	QPQER
1354.	PQERI
1355.	QERIP
1356.	ERIPN
1357.	RIPNS
1358.	IPNSQ
1359.	PNSQQ
1360.	NSQQV
1361.	SQQVE
1362.	QQVEG
1363.	QVEGI
1364.	VEGIL
1365.	EGILR
1366.	GILRD
1367.	ILRDM
1368.	LRDML
1369.	RDMLT
1370.	DMLTN
1371.	MLTNG
1372.	LTNGS
1373.	TNGSQ
1374.	NGSQT
1375.	GSQTF
1376.	SQTFR
1377.	QTFRD
1378.	TFRDL
1379.	FRDLM
1380.	RDLMR
1381.	DLMRR
1382.	LMRRW
1383.	MRRWN
1384.	RRWNR
1385.	RWNRE
1386.	WNREV
1387.	NREVD
1388.	REVDR
1389.	EVDRE

5-mer peptides of CT147 (hypothetical protein,

1449 amino acids), ACCESSION# NP 219650.1, GI:

15604866 (SEQ ID NOS: 1390-2834)

1390.	MANPS
1391.	ANPST
1392.	NPSTP
1393.	PSTPS
1394.	STPSF
1395.	TPSFN
1396.	PSFNH
1397.	SFNHS
1398.	FNHSD
1399.	NHSDL
1400.	HSDLS
1401.	SDLSL
1402.	DLSLQ
1403.	LSLQG
1404.	SLQGR
1405.	LQGRL
1406.	QGRLR
1407.	GRLRA
1408.	RLRAS
1409.	LRASS
1410.	RASSQ
1411.	ASSQQ
1412.	SSQQC
1413.	SQQCT
1414.	QQCTQ
1415.	QCTQA
1416.	CTQAG
1417.	TQAGQ
1418.	QAGQG
1419.	AGQGD
1420.	GQGDP
1421.	QGDPQ
1422.	GDPQP
1423.	DPQPL
1424.	PQPLS
1425.	QPLSP
1426.	PLSPE
1427.	LSPES
1428.	SPESR
1429.	PESRG
1430.	ESRGL
1431.	SRGLT
1432.	RGLTS
1433.	GLTSN
1434.	LTSNF
1435.	TSNFS
1436.	SNFST
1437.	NFSTR
1438.	FSTRR
1439.	STRRD
1440.	TRRDL
1441.	RRDLI
1442.	RDLID
1443.	DLIDV
1444.	LIDVV
1445.	IDVVE
1446.	DVVEE
1447.	VVEES
1448.	VEESI
1449.	EESIE
1450.	ESIET
1451.	SIETA
1452.	IETAK
1453.	ETAKG
1454.	TAKGS
1455.	AKGSE
1456.	KGSEL
1457.	GSELK
1458.	SELKK
1459.	ELKKL
1460.	LKKLR
1461.	KKLRI
1462.	KLRIY
1463.	LRIYE
1464.	RIYEI
1465.	IYEIA
1466.	YEIAL
1467.	EIALK
1468.	IALKI
1469.	ALKIL
1470.	LKILT
1471.	KILTI
1472.	ILTII
1473.	LTIIG
1474.	TIIGA
1475.	IIGAA
1476.	IGAAI
1477.	GAAIL
1478.	AAILF
1479.	AILFA
1480.	ILFAV
1481.	LFAVP
1482.	FAVPL
1483.	AVPLC
1484.	VPLCM
1485.	PLCML
1486.	LCMLL
1487.	CMLLG
1488.	MLLGV
1489.	LLGVP
1490.	LGVPL
1491.	GVPLW
1492.	VPLWI
1493.	PLWIP
1494.	LWIPI
1495.	WIPIV
1496.	IPIVT
1497.	PIVTC
1498.	IVTCI
1499.	VTCIG
1500.	TCIGV
1501.	CIGVG
1502.	IGVGI
1503.	GVGIA
1504.	VGIAF
1505.	GIAFS
1506.	IAFSI
1507.	AFSIA
1508.	FSIAK
1509.	SIAKG
1510.	IAKGC
1511.	AKGCL
1512.	KGCLQ
1513.	GCLQK
1514.	CLQKR
1515.	LQKRC
1516.	QKRCQ
1517.	KRCQQ
1518.	RCQQI
1519.	CQQIR
1520.	QQIRE
1521.	QIREE
1522.	IREEY
1523.	REEYR
1524.	EEYRA
1525.	EYRAL
1526.	YRALH
1527.	RALHL
1528.	ALHLY
1529.	LHLYH
1530.	HLYHR
1531.	LYHRY
1532.	YHRYL
1533.	HRYLL
1534.	RYLLS
1535.	YLLSN
1536.	LLSNK
1537.	LSNKD
1538.	SNKDS
1539.	NKDSI
1540.	KDSID
1541.	DSIDG
1542.	SIDGT
1543.	IDGTL
1544.	DGTLL
1545.	GTLLS
1546.	TLLSR
1547.	LLSRF
1548.	LSRFD
1549.	SRFDI
1550.	RFDIR
1551.	FDIRF
1552.	DIRFR
1553.	IRFRK
1554.	RFRKA
1555.	FRKAE
1556.	RKAEE
1557.	KAEEK
1558.	AEEKL
1559.	EEKLH
1560.	EKLHG
1561.	KLHGL
1562.	LHGLD
1563.	HGLDL
1564.	GLDLD
1565.	LDLDK
1566.	DLDKR
1567.	LDKRE
1568.	DKREA
1569.	KREAN
1570.	REANH
1571.	EANHP
1572.	ANHPL
1573.	NHPLE
1574.	HPLEA
1575.	PLEAD
1576.	LEADK
1577.	EADKR
1578.	ADKRY
1579.	DKRYD
1580.	KRYDF
1581.	RYDFA
1582.	YDFAG
1583.	DFAGL
1584.	FAGLA
1585.	AGLAH
1586.	GLAHQ
1587.	LAHQR
1588.	AHQRY
1589.	HQRYQ
1590.	QRYQV
1591.	RYQVD
1592.	YQVDA
1593.	QVDAA
1594.	VDAAL
1595.	DAALG
1596.	AALGI
1597.	ALGIS
1598.	LGISS
1599.	GISSS
1600.	ISSSQ
1601.	SSSQD
1602.	SSQDA
1603.	SQDAF
1604.	QDAFW
1605.	DAFWR
1606.	AFWRG
1607.	FWRGV
1608.	WRGVA
1609.	RGVAQ
1610.	GVAQQ
1611.	VAQQV
1612.	AQQVK
1613.	QQVKS
1614.	QVKSV
1615.	VKSVK
1616.	KSVKD
1617.	SVKDD
1618.	VKDDV
1619.	KDDVV
1620.	DDVVL
1621.	DVVLG
1622.	VVLGD
1623.	VLGDK
1624.	LGDKA
1625.	GDKAS
1626.	DKAST
1627.	KASTD
1628.	ASTDL
1629.	STDLY
1630.	TDLYP
1631.	DLYPI
1632.	LYPIA
1633.	YPIAQ
1634.	PIAQQ
1635.	IAQQA
1636.	AQQAL
1637.	QQALQ
1638.	QALQA
1639.	ALQAA
1640.	LQAAG
1641.	QAAGV
1642.	AAGVG
1643.	AGVGF
1644.	GVGFS
1645.	VGFSG
1646.	GFSGA
1647.	FSGAA
1648.	SGAAG
1649.	GAAGK
1650.	AAGKE
1651.	AGKES
1652.	GKESL
1653.	KESLL
1654.	ESLLD
1655.	SLLDL
1656.	LLDLA
1657.	LDLAK
1658.	DLAKS
1659.	LAKSL
1660.	AKSLS
1661.	KSLSS
1662.	SLSSL
1663.	LSSLF
1664.	SSLFA
1665.	SLFAW
1666.	LFAWG
1667.	FAWGS
1668.	AWGSQ
1669.	WGSQV
1670.	GSQVG
1671.	SQVGK
1672.	QVGKD
1673.	VGKDS
1674.	GKDSH
1675.	KDSHE
1676.	DSHEA
1677.	SHEAL
1678.	HEALQ
1679.	EALQQ
1680.	ALQQY
1681.	LQQYQ
1682.	QQYQM
1683.	QYQMR
1684.	YQMRF
1685.	QMRFL
1686.	MRFLS
1687.	RFLSS
1688.	FLSSP
1689.	LSSPI
1690.	SSPIL
1691.	SPILA
1692.	PILAT
1693.	ILATW
1694.	LATWC
1695.	ATWCG
1696.	TWCGA
1697.	WCGAG
1698.	CGAGF
1699.	GAGFS
1700.	AGFSA
1701.	GFSAS
1702.	FSASA
1703.	SASAQ
1704.	ASAQD
1705.	SAQDF
1706.	AQDFV
1707.	QDFVL
1708.	DFVLK
1709.	FVLKG
1710.	VLKGE
1711.	LKGEN
1712.	KGENI
1713.	GENIL
1714.	ENILD
1715.	NILDI
1716.	ILDIA
1717.	LDIAS
1718.	DIASE
1719.	IASEN
1720.	ASENH
1721.	SENHT
1722.	ENHTK
1723.	NHTKM
1724.	HTKMQ
1725.	TKMQN
1726.	KMQNA
1727.	MQNAI
1728.	QNAIK
1729.	NAIKR
1730.	AIKRV
1731.	IKRVQ
1732.	KRVQL
1733.	RVQLV
1734.	VQLVS
1735.	QLVSV
1736.	LVSVL
1737.	VSVLG
1738.	SVLGK
1739.	VLGKM
1740.	LGKMR
1741.	GKMRN
1742.	KMRNW
1743.	MRNWK
1744.	RNWKE
1745.	NWKEK
1746.	WKEKI
1747.	KEKID
1748.	EKIDT
1749.	KIDTL
1750.	IDTLI
1751.	DTLIQ
1752.	TLIQN
1753.	LIQNK
1754.	IQNKN
1755.	QNKNL
1756.	NKNLD
1757.	KNLDQ
1758.	NLDQD
1759.	LDQDS
1760.	DQDSL
1761.	QDSLR
1762.	DSLRK
1763.	SLRKL
1764.	LRKLY
1765.	RKLYQ
1766.	KLYQD
1767.	LYQDI
1768.	YQDIE
1769.	QDIEK
1770.	DIEKA
1771.	IEKAM
1772.	EKAMH
1773.	KAMHK
1774.	AMHKV
1775.	MHKVC
1776.	HKVCI
1777.	KVCIE
1778.	VCIED
1779.	CIEDG
1780.	IEDGV
1781.	EDGVS
1782.	DGVST
1783.	GVSTS
1784.	VSTSI
1785.	STSIQ
1786.	TSIQT
1787.	SIQTQ
1788.	IQTQV
1789.	QTQVR
1790.	TQVRK
1791.	QVRKV
1792.	VRKVT
1793.	RKVTQ
1794.	KVTQK
1795.	VTQKY
1796.	TQKYL
1797.	QKYLR
1798.	KYLRQ
1799.	YLRQD
1800.	LRQDL
1801.	RQDLQ
1802.	QDLQE
1803.	DLQEL
1804.	LQELL
1805.	QELLN
1806.	ELLNK
1807.	LLNKK
1808.	LNKKA
1809.	NKKAP
1810.	KKAPL
1811.	KAPLN
1812.	APLNE
1813.	PLNES
1814.	LNESD
1815.	NESDL
1816.	ESDLS
1817.	SDLSK
1818.	DLSKM
1819.	LSKMQ
1820.	SKMQK
1821.	KMQKG
1822.	MQKGI
1823.	QKGIS
1824.	KGISS
1825.	GISSC
1826.	ISSCA
1827.	SSCAN
1828.	SCANL
1829.	CANLV
1830.	ANLVV
1831.	NLVVT
1832.	LVVTL
1833.	VVTLL
1834.	VTLLE
1835.	TLLES
1836.	LLESQ
1837.	LESQL
1838.	ESQLG
1839.	SQLGT
1840.	QLGTS
1841.	LGTSG
1842.	GTSGQ
1843.	TSGQT
1844.	SGQTP
1845.	GQTPI
1846.	QTPIK
1847.	TPIKE
1848.	PIKEV
1849.	IKEVE
1850.	KEVEE
1851.	EVEES
1852.	VEESI
1853.	EESIY
1854.	ESIYR
1855.	SIYRD
1856.	IYRDL
1857.	YRDLI
1858.	RDLIA
1859.	DLIAT
1860.	LIATI
1861.	IATIL
1862.	ATILQ
1863.	TILQM
1864.	ILQMG
1865.	LQMGS
1866.	QMGSA
1867.	MGSAA
1868.	GSAAG
1869.	SAAGG
1870.	AAGGV
1871.	AGGVT
1872.	GGVTP
1873.	GVTPL
1874.	VTPLV
1875.	TPLVD
1876.	PLVDG
1877.	LVDGV
1878.	VDGVH
1879.	DGVHK
1880.	GVHKA
1881.	VHKAI
1882.	HKAIR
1883.	KAIRE
1884.	AIREG
1885.	IREGK
1886.	REGKA
1887.	EGKAL
1888.	GKALR
1889.	KALRS
1890.	ALRSE
1891.	LRSEL
1892.	RSELS
1893.	SELSR
1894.	ELSRA
1895.	LSRAM
1896.	SRAMS
1897.	RAMSL
1898.	AMSLH
1899.	MSLHP
1900.	SLHPR
1901.	LHPRQ
1902.	HPRQS
1903.	PRQSF
1904.	RQSFL
1905.	QSFLG
1906.	SFLGV
1907.	FLGVQ
1908.	LGVQS
1909.	GVQSA
1910.	VQSAV
1911.	QSAVE
1912.	SAVEK
1913.	AVEKL
1914.	VEKLQ
1915.	EKLQA
1916.	KLQAF
1917.	LQAFI
1918.	QAFIR
1919.	AFIRD
1920.	FIRDP
1921.	IRDPK
1922.	RDPKW
1923.	DPKWG
1924.	PKWGA
1925.	KWGAS
1926.	WGASA
1927.	GASAV
1928.	ASAVH
1929.	SAVHT
1930.	AVHTS
1931.	VHTSA
1932.	HTSAE
1933.	TSAEE
1934.	SAEET
1935.	AEETL
1936.	EETLA
1937.	ETLAQ
1938.	TLAQK
1939.	LAQKQ
1940.	AQKQK
1941.	QKQKF
1942.	KQKFV
1943.	QKFVS
1944.	KFVSD
1945.	FVSDL
1946.	VSDLT
1947.	SDLTR
1948.	DLTRI
1949.	LTRIQ
1950.	TRIQT
1951.	RIQTS
1952.	IQTSL
1953.	QTSLA
1954.	TSLAD
1955.	SLADW
1956.	LADWR
1957.	ADWRE
1958.	DWRER
1959.	WRERY
1960.	RERYG
1961.	ERYGL
1962.	RYGLF
1963.	YGLFE
1964.	GLFEE
1965.	LFEET
1966.	FEETK
1967.	EETKL
1968.	ETKLN
1969.	TKLNH
1970.	KLNHI
1971.	LNHIV
1972.	NHIVS
1973.	HIVST
1974.	IVSTD
1975.	VSTDF
1976.	STDFV
1977.	TDFVS
1978.	DFVSR
1979.	FVSRT
1980.	VSRTE
1981.	SRTEA
1982.	RTEAF
1983.	TEAFL
1984.	EAFLD
1985.	AFLDT
1986.	FLDTL
1987.	LDTLK
1988.	DTLKN
1989.	TLKNV
1990.	LKNVA
1991.	KNVAE
1992.	NVAEA
1993.	VAEAC
1994.	AEACS
1995.	EACSL
1996.	ACSLE
1997.	CSLEQ
1998.	SLEQA
1999.	LEQAV
2000.	EQAVA
2001.	QAVAE
2002.	AVAEL
2003.	VAELK
2004.	AELKD
2005.	ELKDC
2006.	LKDCE
2007.	KDCED
2008.	DCEDA
2009.	CEDAM
2010.	EDAMK
2011.	DAMKA
2012.	AMKAD
2013.	MKADL
2014.	KADLT
2015.	ADLTH
2016.	DLTHV
2017.	LTHVE
2018.	THVEQ
2019.	HVEQK
2020.	VEQKM
2021.	EQKMN
2022.	QKMNP
2023.	KMNPT
2024.	MNPTE
2025.	NPTEI
2026.	PTEIE
2027.	TEIES
2028.	EIESA
2029.	IESAR
2030.	ESARE
2031.	SAREE
2032.	AREEF
2033.	REEFK
2034.	EEFKR
2035.	EFKRL
2036.	FKRLM
2037.	KRLME
2038.	RLMEE
2039.	LMEEL
2040.	MEELA
2041.	EELAG
2042.	ELAGI
2043.	LAGIQ
2044.	AGIQE
2045.	GIQEQ
2046.	IQEQL
2047.	QEQLE
2048.	EQLEQ
2049.	QLEQI
2050.	LEQIA
2051.	EQIAQ
2052.	QIAQP
2053.	IAQPI
2054.	AQPIY
2055.	QPIYE
2056.	PIYEE
2057.	IYEEG
2058.	YEEGV
2059.	EEGVS
2060.	EGVSG
2061.	GVSGE
2062.	VSGER
2063.	SGERL
2064.	GERLL
2065.	ERLLL
2066.	RLLLN
2067.	LLLNT
2068.	LLNTV
2069.	LNTVF
2070.	NTVFF
2071.	TVFFH
2072.	VFFHP
2073.	FFHPE
2074.	FHPEV
2075.	HPEVL
2076.	PEVLR
2077.	EVLRK
2078.	VLRKK
2079.	LRKKV
2080.	RKKVQ
2081.	KKVQA
2082.	KVQAK
2083.	VQAKE
2084.	QAKEA
2085.	AKEAS
2086.	KEASL
2087.	EASLE
2088.	ASLEA
2089.	SLEAL
2090.	LEALT
2091.	EALTK
2092.	ALTKG
2093.	LTKGE
2094.	TKGEQ
2095.	KGEQP
2096.	GEQPS
2097.	EQPSP
2098.	QPSPT
2099.	PSPTK
2100.	SPTKK
2101.	PTKKK
2102.	TKKKT
2103.	KKKTL
2104.	KKTLK
2105.	KTLKQ
2106.	TLKQL
2107.	LKQLS
2108.	KQLSE
2109.	QLSEG
2110.	LSEGC
2111.	SEGCE
2112.	EGCEY
2113.	GCEYF
2114.	CEYFS
2115.	EYFSS
2116.	YFSSL
2117.	FSSLV
2118.	SSLVS
2119.	SLVSK
2120.	LVSKI
2121.	VSKIN
2122.	SKINA
2123.	KINAL
2124.	INALK
2125.	NALKT
2126.	ALKTI
2127.	LKTIL
2128.	KTILE
2129.	TILEG
2130.	ILEGS
2131.	LEGSR
2132.	EGSRG
2133.	GSRGK
2134.	SRGKK
2135.	RGKKI
2136.	GKKIA
2137.	KKIAS
2138.	KIASQ
2139.	IASQD
2140.	ASQDI
2141.	SQDIR
2142.	QDIRQ
2143.	DIRQL
2144.	IRQLI
2145.	RQLIG
2146.	QLIGL
2147.	LIGLT
2148.	IGLTD
2149.	GLTDE
2150.	LTDEL
2151.	TDELA
2152.	DELAL
2153.	ELALE
2154.	LALEL
2155.	ALELS
2156.	LELSS
2157.	ELSSF
2158.	LSSFQ
2159.	SSFQQ
2160.	SFQQD
2161.	FQQDS
2162.	QQDSL
2163.	QDSLE
2164.	DSLES
2165.	SLESL
2166.	LESLL
2167.	ESLLY
2168.	SLLYG
2169.	LLYGL
2170.	LYGLE
2171.	YGLEG
2172.	GLEGL
2173.	LEGLS
2174.	EGLSI
2175.	GLSIP
2176.	LSIPA
2177.	SIPAA
2178.	IPAAS
2179.	PAASI
2180.	AASIE
2181.	ASIEQ
2182.	SIEQK
2183.	IEQKK
2184.	EQKKG
2185.	QKKGS
2186.	KKGSP
2187.	KGSPK
2188.	GSPKS
2189.	SPKSS
2190.	PKSSS
2191.	KSSSI
2192.	SSSIA
2193.	SSIAE
2194.	SIAEK
2195.	IAEKV
2196.	AEKVV
2197.	EKVVY
2198.	KVVYA
2199.	VVYAS
2200.	VYASH
2201.	YASHQ
2202.	ASHQR
2203.	SHQRV
2204.	HQRVH
2205.	QRVHN
2206.	RVHNG
2207.	VHNGV
2208.	HNGVK
2209.	NGVKA
2210.	GVKAK
2211.	VKAKV
2212.	KAKVN
2213.	AKVNR
2214.	KVNRT
2215.	VNRTL
2216.	NRTLE
2217.	RTLEA
2218.	TLEAF
2219.	LEAFS
2220.	EAFSQ
2221.	AFSQL
2222.	FSQLI
2223.	SQLIK
2224.	QLIKG
2225.	LIKGL
2226.	IKGLR
2227.	KGLRG
2228.	GLRGS
2229.	LRGSL
2230.	RGSLR
2231.	GSLRN
2232.	SLRNA
2233.	LRNAM
2234.	RNAMI
2235.	NAMIT
2236.	AMITK
2237.	MITKA
2238.	ITKAV
2239.	TKAVV
2240.	KAVVA
2241.	AVVAA
2242.	VVAAV
2243.	VAAVL
2244.	AAVLS
2245.	AVLSV
2246.	VLSVA
2247.	LSVAF
2248.	SVAFS
2249.	VAFSC
2250.	AFSCL
2251.	FSCLA
2252.	SCLAI
2253.	CLAIA
2254.	LAIAL
2255.	AIALF
2256.	IALFS
2257.	ALFSV
2258.	LFSVQ
2259.	FSVQL
2260.	SVQLT
2261.	VQLTW
2262.	QLTWL
2263.	LTWLP
2264.	TWLPI
2265.	WLPIM
2266.	LPIML
2267.	PIMLC
2268.	IMLCV
2269.	MLCVL
2270.	LCVLA
2271.	CVLAL
2272.	VLALV
2273.	LALVL
2274.	ALVLE
2275.	LVLEA
2276.	VLEAI
2277.	LEAIP
2278.	EAIPS
2279.	AIPSA
2280.	IPSAL
2281.	PSALS
2282.	SALSI
2283.	ALSIW
2284.	LSIWV
2285.	SIWVE
2286.	IWVEK
2287.	WVEKR
2288.	VEKRN
2289.	EKRNW
2290.	KRNWK
2291.	RNWKY
2292.	NWKYE
2293.	WKYEV
2294.	KYEVA
2295.	YEVAS
2296.	EVASL
2297.	VASLA
2298.	ASLAK
2299.	SLAKQ
2300.	LAKQL
2301.	AKQLV
2302.	KQLVS
2303.	QLVSD
2304.	LVSDG
2305.	VSDGR
2306.	SDGRK
2307.	DGRKL
2308.	GRKLP
2309.	RKLPY
2310.	KLPYP
2311.	LPYPD
2312.	PYPDL
2313.	YPDLG
2314.	PDLGD
2315.	DLGDQ
2316.	LGDQN
2317.	GDQNI
2318.	DQNIK
2319.	QNIKH
2320.	NIKHL
2321.	IKHLE
2322.	KHLEK
2323.	HLEKI
2324.	LEKIR
2325.	EKIRD
2326.	KIRDV
2327.	IRDVY
2328.	RDVYG
2329.	DVYGL
2330.	VYGLD
2331.	YGLDG
2332.	GLDGV
2333.	LDGVA
2334.	DGVAE
2335.	GVAEL
2336.	VAELR
2337.	AELRV
2338.	ELRVA
2339.	LRVAE
2340.	RVAEA
2341.	VAEAA
2342.	AEAAL
2343.	EAALL
2344.	AALLG
2345.	ALLGV
2346.	LLGVQ
2347.	LGVQK
2348.	GVQKL
2349.	VQKLP
2350.	QKLPE
2351.	KLPEE
2352.	LPEEQ
2353.	PEEQK
2354.	EEQKQ
2355.	EQKQE
2356.	QKQES
2357.	KQESL
2358.	QESLK
2359.	ESLKS
2360.	SLKSA
2361.	LKSAV
2362.	KSAVK
2363.	SAVKA
2364.	AVKAL
2365.	VKALR
2366.	KALRA
2367.	ALRAD
2368.	LRADA
2369.	RADAK
2370.	ADAKV
2371.	DAKVL
2372.	AKVLN
2373.	KVLNK
2374.	VLNKK
2375.	LNKKF
2376.	NKKFK
2377.	KKFKK
2378.	KFKKL
2379.	FKKLP
2380.	KKLPE
2381.	KLPES
2382.	LPESY
2383.	PESYQ
2384.	ESYQP
2385.	SYQPQ
2386.	YQPQH
2387.	QPQHS
2388.	PQHSE
2389.	QHSEV
2390.	HSEVT
2391.	SEVTG
2392.	EVTGV
2393.	VTGVQ
2394.	TGVQG
2395.	GVQGV
2396.	VQGVT
2397.	QGVTE
2398.	GVTEQ
2399.	VTEQE
2400.	TEQES
2401.	EQESR
2402.	QESRD
2403.	ESRDD
2404.	SRDDV
2405.	RDDVL
2406.	DDVLV
2407.	DVLVA
2408.	VLVAQ
2409.	LVAQD
2410.	VAQDM
2411.	AQDMA
2412.	QDMAA
2413.	DMAAI
2414.	MAAIE
2415.	AAIEE
2416.	AIEEL
2417.	IEELQ
2418.	EELQD
2419.	ELQDQ
2420.	LQDQY
2421.	QDQYH
2422.	DQYHA
2423.	QYHAA
2424.	YHAAC
2425.	HAACL
2426.	AACLQ
2427.	ACLQF
2428.	CLQFE
2429.	LQFES
2430.	QFESV
2431.	FESVS
2432.	ESVST
2433.	SVSTR
2434.	VSTRF
2435.	STRFL
2436.	TRFLA
2437.	RFLAE
2438.	FLAEQ
2439.	LAEQR
2440.	AEQRK
2441.	EQRKA
2442.	QRKAK
2443.	RKAKF
2444.	KAKFL
2445.	AKFLE
2446.	KFLEE
2447.	FLEEL
2448.	LEELL
2449.	EELLV
2450.	ELLVQ
2451.	LLVQK
2452.	LVQKR
2453.	VQKRR
2454.	QKRRD
2455.	KRRDV
2456.	RRDVS
2457.	RDVSH
2458.	DVSHL
2459.	VSHLS
2460.	SHLSH
2461.	HLSHQ
2462.	LSHQE
2463.	SHQEA
2464.	HQEAH
2465.	QEAHY
2466.	EAHYT
2467.	AHYTQ
2468.	HYTQV
2469.	YTQVV
2470.	TQVVS
2471.	QVVSH
2472.	VVSHL
2473.	VSHLK
2474.	SHLKE
2475.	HLKEL
2476.	LKELI
2477.	KELIS
2478.	ELISM
2479.	LISMR
2480.	ISMRK
2481.	SMRKG
2482.	MRKGA
2483.	RKGAS
2484.	KGAST
2485.	GASTQ
2486.	ASTQH
2487.	STQHA
2488.	TQHAS
2489.	QHASK
2490.	HASKE
2491.	ASKEE
2492.	SKEET
2493.	KEEIS
2494.	EEIST
2495.	EISTK
2496.	ISTKM
2497.	STKMR
2498.	TKMRE
2499.	KMREL
2500.	MRELL
2501.	RELLS
2502.	ELLSL
2503.	LLSLD
2504.	LSLDD
2505.	SLDDQ
2506.	LDDQL
2507.	DDQLL
2508.	DQLLK
2509.	QLLKA
2510.	LLKAH
2511.	LKAHT
2512.	KAHTA
2513.	AHTAQ
2514.	HTAQD
2515.	TAQDV
2516.	AQDVN
2517.	QDVNR
2518.	DVNRD
2519.	VNRDN
2520.	NRDNS
2521.	RDNSI
2522.	DNSIN
2523.	NSING
2524.	SINGQ
2525.	INGQL
2526.	NGQLQ
2527.	GQLQQ
2528.	QLQQQ
2529.	LQQQF
2530.	QQQFK
2531.	QQFKK
2532.	QFKKL
2533.	FKKLS
2534.	KKLSE
2535.	KLSEE
2536.	LSEEG
2537.	SEEGS
2538.	EEGSL
2539.	EGSLQ
2540.	GSLQK
2541.	SLQKV
2542.	LQKVK
2543.	QKVKA
2544.	KVKAL
2545.	VKALL
2546.	KALLE
2547.	ALLEL
2548.	LLELN
2549.	LELNM
2550.	ELNMC
2551.	LNMCL
2552.	NMCLG
2553.	MCLGN
2554.	CLGNA
2555.	LGNAG
2556.	GNAGQ
2557.	NAGQT
2558.	AGQTL
2559.	GQTLY
2560.	QTLYH
2561.	TLYHS
2562.	LYHSR
2563.	YHSRL
2564.	HSRLK
2565.	SRLKR
2566.	RLKRE
2567.	LKREV
2568.	KREVF
2569.	REVFE
2570.	EVFEA
2571.	VFEAS
2572.	FEASL
2573.	EASLS
2574.	ASLSG
2575.	SLSGT
2576.	LSGTS
2577.	SGTSR
2578.	GTSRQ
2579.	TSRQL
2580.	SRQLL
2581.	RQLLQ
2582.	QLLQY
2583.	LLQYG
2584.	LQYGE
2585.	QYGED
2586.	YGEDL
2587.	GEDLF
2588.	EDLFA
2589.	DLFAS
2590.	LFASY
2591.	FASYD
2592.	ASYDG
2593.	SYDGS
2594.	YDGSD
2595.	DGSDR
2596.	GSDRS
2597.	SDRSA
2598.	DRSAL
2599.	RSALL
2600.	SALLR
2601.	ALLRF
2602.	LLRFV
2603.	LRFVL
2604.	RFVLG
2605.	FVLGS
2606.	VLGSG
2607.	LGSGY
2608.	GSGYE
2609.	SGYEM
2610.	GYEMI
2611.	YEMIS
2612.	EMISE
2613.	MISEA
2614.	ISEAS
2615.	SEASS
2616.	EASSE
2617.	ASSEL
2618.	SSELK
2619.	SELKS
2620.	ELKSL
2621.	LKSLR
2622.	KSLRK
2623.	SLRKR
2624.	LRKRW
2625.	RKRWK
2626.	KRWKR
2627.	RWKRS
2628.	WKRSA
2629.	KRSAS
2630.	RSASQ
2631.	SASQA
2632.	ASQAA
2633.	SQAAI
2634.	QAAIA
2635.	AAIAP
2636.	AIAPE
2637.	IAPED
2638.	APEDY
2639.	PEDYE
2640.	EDYEK
2641.	DYEKV
2642.	YEKVC
2643.	EKVCR
2644.	KVCRV
2645.	VCRVL
2646.	CRVLE
2647.	RVLER
2648.	VLERF
2649.	LERFL
2650.	ERFLK
2651.	RFLKA
2652.	FLKAR
2653.	LKARD
2654.	KARDS
2655.	ARDSL
2656.	RDSLR
2657.	DSLRP
2658.	SLRPK
2659.	LRPKL
2660.	RPKLG
2661.	PKLGL
2662.	KLGLP
2663.	LGLPL
2664.	GLPLG
2665.	LPLGK
2666.	PLGKS
2667.	LGKSS
2668.	GKSSD
2669.	KSSDA
2670.	SSDAT
2671.	SDATV
2672.	DATVG
2673.	ATVGL
2674.	TVGLQ
2675.	VGLQH
2676.	GLQHQ
2677.	LQHQI
2678.	QHQIR
2679.	HQIRD
2680.	QIRDN
2681.	IRDNQ
2682.	RDNQR
2683.	DNQRV
2684.	NQRVK
2685.	QRVKA
2686.	RVKAR
2687.	VKARV
2688.	KARVT
2689.	ARVTA
2690.	RVTAC
2691.	VTACY
2692.	TACYQ
2693.	ACYQE
2694.	CYQES
2695.	YQESC
2696.	QESCR
2697.	ESCRN
2698.	SCRNV
2699.	CRNVL
2700.	RNVLQ
2701.	NVLQH
2702.	VLQHL
2703.	LQHLE
2704.	QHLED
2705.	HLEDW
2706.	LEDWV
2707.	EDWVR
2708.	DWVRK
2709.	WVRKT
2710.	VRKTR
2711.	RKTRQ
2712.	KTRQE
2713.	TRQES
2714.	RQESA
2715.	QESAE
2716.	ESAEC
2717.	SAECQ
2718.	AECQK
2719.	ECQKV
2720.	CQKVE
2721.	QKVET
2722.	KVETK
2723.	VETKI
2724.	ETKIR
2725.	TKIRE
2726.	KIREF
2727.	IREFC
2728.	REFCQ
2729.	EFCQK
2730.	FCQKA
2731.	CQKAG
2732.	QKAGS
2733.	KAGSK
2734.	AGSKE
2735.	GSKEN
2736.	SKENL
2737.	KENLA
2738.	ENLAE
2739.	NLAES
2740.	LAEST
2741.	AESTE
2742.	ESTEM
2743.	STEML
2744.	TEMLF
2745.	EMLFS
2746.	MLFSS
2747.	LFSSL
2748.	FSSLE
2749.	SSLEE
2750.	SLEED
2751.	LEEDL
2752.	EEDLN
2753.	EDLNK
2754.	DLNKI
2755.	LNKIP
2756.	NKIPL
2757.	KIPLD
2758.	IPLDV
2759.	PLDVL
2760.	LDVLR
2761.	DVLRA
2762.	VLRAI
2763.	LRAIL
2764.	RAILR
2765.	AILRS
2766.	ILRSL
2767.	LRSLS
2768.	RSLSS
2769.	SLSSK
2770.	LSSKV
2771.	SSKVL
2772.	SKVLH
2773.	KVLHI
2774.	VLHIR
2775.	LHIRD
2776.	HIRDQ
2777.	IRDQK
2778.	RDQKL
2779.	DQKLE
2780.	QKLEL
2781.	KLELE
2782.	LELEK
2783.	ELEKL
2784.	LEKLE
2785.	EKLEE
2786.	KLEEQ
2787.	LEEQF
2788.	EEQFA
2789.	EQFAK
2790.	QFAKT
2791.	FAKTN
2792.	AKTNA
2793.	KTNAI
2794.	TNAIV
2795.	NAIVK
2796.	AIVKA
2797.	IVKAK
2798.	VKAKE
2799.	KAKEA
2800.	AKEAE
2801.	KEAEF
2802.	EAEFE
2803.	AEFEK
2804.	EFEKN
2805.	FEKNG
2806.	EKNGE
2807.	KNGEV
2808.	NGEVW
2809.	GEVWH
2810.	EVWHN
2811.	VWHNQ
2812.	WHNQY
2813.	HNQYQ
2814.	NQYQM
2815.	QYQML
2816.	YQMLK
2817.	QMLKS
2818.	MLKSQ
2819.	LKSQM
2820.	KSQME
2821.	SQMEK
2822.	QMEKL
2823.	MEKLE
2824.	EKLES
2825.	KLESQ
2826.	LESQK
2827.	ESQKR
2828.	SQKRR
2829.	QKRRL
2830.	KRRLT
2831.	RRLTD
2832.	RLTDK
2833.	LTDKK
2834.	TDKKE

5-mer peptides of HSP60 (CT110, 544 amino acids),

ACCESSION# AAS19616.1, GI: 42541742 (SEQ ID NOS:

2835-3374)

2835.	MVAKN
2836.	VAKNI
2837.	AKNIK
2838.	KNIKY
2839.	NIKYN
2840.	IKYNE
2841.	KYNEE
2842.	YNEEA
2843.	NEEAR
2844.	EEARK
2845.	EARKK
2846.	ARKKI
2847.	RKKIQ
2848.	KKIQK
2849.	KIQKG
2850.	IQKGV
2851.	QKGVK
2852.	KGVKT
2853.	GVKTL
2854.	VKTLA
2855.	KTLAE
2856.	TLAEA
2857.	LAEAV
2858.	AEAVK
2859.	EAVKV
2860.	AVKVT
2861.	VKVTL
2862.	KVTLG
2863.	VTLGP
2864.	TLGPK
2865.	LGPKG
2866.	GPKGR
2867.	PKGRH
2868.	KGRHV
2869.	GRHVV
2870.	RHVVI
2871.	HVVID
2872.	VVIDK
2873.	VIDKS
2874.	IDKSF
2875.	DKSFG
2876.	KSFGS
2877.	SFGSP
2878.	FGSPQ
2879.	GSPQV
2880.	SPQVT
2881.	PQVTK
2882.	QVTKD
2883.	VTKDG
2884.	TKDGV
2885.	KDGVT
2886.	DGVTV
2887.	GVTVA
2888.	VTVAK
2889.	TVAKE
2890.	VAKEV
2891.	AKEVE
2892.	KEVEL
2893.	EVELA
2894.	VELAD
2895.	ELADK
2896.	LADKH
2897.	ADKHE
2898.	DKHEN
2899.	KHENM
2900.	HENMG
2901.	ENMGA
2902.	NMGAQ
2903.	MGAQM
2904.	GAQMV
2905.	AQMVK
2906.	QMVKE
2907.	MVKEV
2908.	VKEVA
2909.	KEVAS
2910.	EVASK
2911.	VASKT
2912.	ASKTA
2913.	SKTAD
2914.	KTADK
2915.	TADKA
2916.	ADKAG
2917.	DKAGD
2918.	KAGDG
2919.	AGDGT
2920.	GDGTT
2921.	DGTTT
2922.	GTTTA
2923.	TTTAT
2924.	TTATV
2925.	TATVL
2926.	ATVLA
2927.	TVLAE
2928.	VLAEA
2929.	LAEAI
2930.	AEAIY
2931.	EAIYT
2932.	AIYTE
2933.	IYTEG
2934.	YTEGL
2935.	TEGLR
2936.	EGLRN
2937.	GLRNV
2938.	LRNVT
2939.	RNVTA
2940.	NVTAG
2941.	VTAGA
2942.	TAGAN
2943.	AGANP
2944.	GANPM
2945.	ANPMD
2946.	NPMDL
2947.	PMDLK
2948.	MDLKR
2949.	DLKRG
2950.	LKRGI
2951.	KRGID
2952.	RGIDK
2953.	GIDKA
2954.	IDKAV
2955.	DKAVK
2956.	KAVKV
2957.	AVKVV
2958.	VKVVV
2959.	KVVVD
2960.	VVVDQ
2961.	VVDQI
2962.	VDQIR
2963.	DQIRK
2964.	QIRKI
2965.	IRKIS
2966.	RKISK
2967.	KISKP
2968.	ISKPV
2969.	SKPVQ
2970.	KPVQH
2971.	PVQHH
2972.	VQHHK
2973.	QHHKE
2974.	HHKEI
2975.	HKEIA
2976.	KEIAQ
2977.	EIAQV
2978.	IAQVA
2979.	AQVAT
2980.	QVATI
2981.	VATIS
2982.	ATISA
2983.	TISAN
2984.	ISANN
2985.	SANND
2986.	ANNDA
2987.	NNDAE
2988.	NDAEI
2989.	DAEIG
2990.	AEIGN
2991.	EIGNL
2992.	IGNLI
2993.	GNLIA
2994.	NLIAE
2995.	LIAEA
2996.	IAEAM
2997.	AEAME
2998.	EAMEK
2999.	AMEKV
3000.	MEKVG
3001.	EKVGK
3002.	KVGKN
3003.	VGKNG
3004.	GKNGS
3005.	KNGSI
3006.	NGSIT
3007.	GSITV
3008.	SITVE
3009.	ITVEE
3010.	TVEEA
3011.	VEEAK
3012.	EEAKG
3013.	EAKGF
3014.	AKGFE
3015.	KGFET
3016.	GFETV
3017.	FETVL
3018.	ETVLD
3019.	TVLDV
3020.	VLDVV
3021.	LDVVE
3022.	DVVEG
3023.	VVEGM
3024.	VEGMN
3025.	EGMNF
3026.	GMNFN
3027.	MNFNR
3028.	NFNRG
3029.	FNRGY
3030.	NRGYL
3031.	RGYLS
3032.	GYLSS
3033.	YLSSY
3034.	LSSYF
3035.	SSYFA
3036.	SYFAT
3037.	YFATN
3038.	FATNP
3039.	ATNPE
3040.	TNPET
3041.	NPETQ
3042.	PETQE
3043.	ETQEC
3044.	TQECV
3045.	QECVL
3046.	ECVLE
3047.	CVLED
3048.	VLEDA
3049.	LEDAL
3050.	EDALV
3051.	DALVL
3052.	ALVLI
3053.	LVLIY
3054.	VLIYD
3055.	LIYDK
3056.	IYDKK
3057.	YDKKI
3058.	DKKIS
3059.	KKISG
3060.	KISGI
3061.	ISGIK
3062.	SGIKD
3063.	GIKDF
3064.	IKDFL
3065.	KDFLP
3066.	DFLPV
3067.	FLPVL
3068.	LPVLQ
3069.	PVLQQ
3070.	VLQQV
3071.	LQQVA
3072.	QQVAE
3073.	QVAES
3074.	VAESG
3075.	AESGR
3076.	ESGRP
3077.	SGRPL
3078.	GRPLL
3079.	RPLLI
3080.	PLLII
3081.	LLIIA
3082.	LIIAE
3083.	IIAED
3084.	IAEDI
3085.	AEDIE
3086.	EDIEG
3087.	DIEGE
3088.	IEGEA
3089.	EGEAL
3090.	GEALA
3091.	EALAT
3092.	ALATL
3093.	LATLV
3094.	ATLVV
3095.	TLVVN
3096.	LVVNR
3097.	VVNRI
3098.	VNRIR
3099.	NRIRG
3100.	RIRGG
3101.	IRGGF
3102.	RGGFR
3103.	GGFRV
3104.	GFRVC
3105.	FRVCA
3106.	RVCAV
3107.	VCAVK
3108.	CAVKA
3109.	AVKAP
3110.	VKAPG
3111.	KAPGF
3112.	APGFG
3113.	PGFGD
3114.	GFGDR
3115.	FGDRR
3116.	GDRRK
3117.	DRRKA
3118.	RRKAM
3119.	RKAML
3120.	KAMLE
3121.	AMLED
3122.	MLEDI
3123.	LEDIA
3124.	EDIAI
3125.	DIAIL
3126.	IAILT
3127.	AILTG
3128.	ILTGG
3129.	LTGGQ
3130.	TGGQL
3131.	GGQLI
3132.	GQLIS
3133.	QLISE
3134.	LISEE
3135.	ISEEL
3136.	SEELG
3137.	EELGM
3138.	ELGMK
3139.	LGMKL
3140.	GMKLE
3141.	MKLEN
3142.	KLENA
3143.	LENAN
3144.	ENANL
3145.	NANLA
3146.	ANLAM
3147.	NLAML
3148.	LAMLG
3149.	AMLGK
3150.	MLGKA
3151.	LGKAK
3152.	GKAKK
3153.	KAKKV
3154.	AKKVI
3155.	KKVIV
3156.	KVIVS
3157.	VIVSK
3158.	IVSKE
3159.	VSKED
3160.	SKEDT
3161.	KEDTT
3162.	EDTTI
3163.	DTTIV
3164.	TTIVE
3165.	TIVEG
3166.	IVEGM
3167.	VEGMG
3168.	EGMGE
3169.	GMGEK
3170.	MGEKE
3171.	GEKEA
3172.	EKEAL
3173.	KEALE
3174.	EALEA
3175.	ALEAR
3176.	LEARC
3177.	EARCE
3178.	ARCES
3179.	RCESI
3180.	CESIK
3181.	ESIKK
3182.	SIKKQ
3183.	IKKQI
3184.	KKQIE
3185.	KQIED
3186.	QIEDS
3187.	IEDSS
3188.	EDSSS
3189.	DSSSD
3190.	SSSDY
3191.	SSDYD
3192.	SDYDK
3193.	DYDKE
3194.	YDKEK
3195.	DKEKL
3196.	KEKLQ
3197.	EKLQE
3198.	KLQER
3199.	LQERL
3200.	QERLA
3201.	ERLAK
3202.	RLAKL
3203.	LAKLS
3204.	AKLSG
3205.	KLSGG
3206.	LSGGV
3207.	SGGVA
3208.	GGVAV
3209.	GVAVI
3210.	VAVIR
3211.	AVIRV
3212.	VIRVG
3213.	IRVGA
3214.	RVGAA
3215.	VGAAT
3216.	GAATE
3217.	AATEI
3218.	ATEIE
3219.	TEIEM
3220.	EIEMK
3221.	IEMKE
3222.	EMKEK
3223.	MKEKK
3224.	KEKKD
3225.	EKKDR
3226.	KKDRV
3227.	KDRVD
3228.	DRVDD
3229.	RVDDA
3230.	VDDAQ
3231.	DDAQH
3232.	DAQHA
3233.	AQHAT
3234.	QHATI
3235.	HATIA
3236.	ATIAA
3237.	TIAAV
3238.	IAAVE
3239.	AAVEE
3240.	AVEEG
3241.	VEEGI
3242.	EEGIL
3243.	EGILP
3244.	GILPG
3245.	ILPGG
3246.	LPGGG
3247.	PGGGT
3248.	GGGTA
3249.	GGTAL
3250.	GTALI
3251.	TALIR
3252.	ALIRC
3253.	LIRCI
3254.	IRCIP
3255.	RCIPT
3256.	CIPTL
3257.	IPTLE
3258.	PTLEA
3259.	TLEAF
3260.	LEAFL
3261.	EAFLP
3262.	AFLPM
3263.	FLPML
3264.	LPMLT
3265.	PMLTN
3266.	MLTNE
3267.	LTNED
3268.	TNEDE
3269.	NEDEQ
3270.	EDEQI
3271.	DEQIG
3272.	EQIGA
3273.	QIGAR
3274.	IGARI
3275.	GARIV
3276.	ARIVL
3277.	RIVLK
3278.	IVLKA
3279.	VLKAL
3280.	LKALS
3281.	KALSA
3282.	ALSAP
3283.	LSAPL
3284.	SAPLK
3285.	APLKQ
3286.	PLKQI
3287.	LKQIA
3288.	KQIAA
3289.	QIAAN
3290.	IAANA
3291.	AANAG
3292.	ANAGK
3293.	NAGKE
3294.	AGKEG
3295.	GKEGA
3296.	KEGAI
3297.	EGAII
3298.	GAIIF
3299.	AIIFQ
3300.	IIFQQ
3301.	IFQQV
3302.	FQQVM
3303.	QQVMS
3304.	QVMSR
3305.	VMSRS
3306.	MSRSA
3307.	SRSAN
3308.	RSANE
3309.	SANEG
3310.	ANEGY
3311.	NEGYD
3312.	EGYDA
3313.	GYDAL
3314.	YDALR
3315.	DALRD
3316.	ALRDA
3317.	LRDAY
3318.	RDAYT
3319.	DAYTD
3320.	AYTDM
3321.	YTDML
3322.	TDMLE
3323.	DMLEA
3324.	MLEAG
3325.	LEAGI
3326.	EAGIL
3327.	AGILD
3328.	GILDP
3329.	ILDPA
3330.	LDPAK
3331.	DPAKV
3332.	PAKVT
3333.	AKVTR
3334.	KVTRS
3335.	VTRSA
3336.	TRSAL
3337.	RSALE
3338.	SALES
3339.	ALESA
3340.	LESAA
3341.	ESAAS
3342.	SAASV
3343.	AASVA
3344.	ASVAG
3345.	SVAGL
3346.	VAGLL
3347.	AGLLL
3348.	GLLLT
3349.	LLLTT
3350.	LLTTE
3351.	LTTEA
3352.	TTEAL
3353.	TEALI
3354.	EALIA
3355.	ALIAE
3356.	LIAEI
3357.	IAEIP
3358.	AEIPE
3359.	EIPEE
3360.	IPEEK
3361.	PEEKP
3362.	EEKPA
3363.	EKPAA
3364.	KPAAA
3365.	PAAAP
3366.	AAAPA
3367.	AAPAM
3368.	APAMP
3369.	PAMPG
3370.	AMPGA
3371.	MPGAG
3372.	PGAGM
3373.	GAGMD
3374.	AGMDY

5-mer peptides of CT376 (malate dehydrogenase, 326

amino acids) ACCESSION# NP 219885.1, GI: 15605100

(SEQ ID NOS: 3375-3696)

3375.	MVSQT
3376.	VSQTV
3377.	SQTVS
3378.	QTVSV
3379.	TVSVA
3380.	VSVAV
3381.	SVAVT
3382.	VAVTG
3383.	AVTGG
3384.	VTGGT
3385.	TGGTG
3386.	GGTGQ
3387.	GTGQI
3388.	TGQIA
3389.	GQIAY
3390.	QIAYS
3391.	IAYSF
3392.	AYSFL
3393.	YSFLF
3394.	SFLFS
3395.	FLFSL
3396.	LFSLA
3397.	FSLAH
3398.	SLAHG
3399.	LAHGD
3400.	AHGDV
3401.	HGDVF
3402.	GDVFG
3403.	DVFGL
3404.	VFGLD
3405.	FGLDC
3406.	GLDCG
3407.	LDCGI
3408.	DCGID
3409.	CGIDL
3410.	GIDLR
3411.	IDLRI
3412.	DLRIY
3413.	LRIYD
3414.	RIYDI
3415.	IYDIP
3416.	YDIPG
3417.	DIPGT
3418.	IPGTE
3419.	PGTER
3420.	GTERA
3421.	TERAL
3422.	ERALS
3423.	RALSG
3424.	ALSGV
3425.	LSGVR
3426.	SGVRM
3427.	GVRME
3428.	VRMEL
3429.	RMELD
3430.	MELDD
3431.	ELDDG
3432.	LDDGA
3433.	DDGAF
3434.	DGAFP
3435.	GAFPL
3436.	AFPLL
3437.	FPLLQ
3438.	PLLQR
3439.	LLQRV
3440.	LQRVQ
3441.	QRVQV
3442.	RVQVT
3443.	VQVTT
3444.	QVTTS
3445.	VTTSL
3446.	TTSLH
3447.	TSLHD
3448.	SLHDA
3449.	LHDAF
3450.	HDAFD
3451.	DAFDG
3452.	AFDGI
3453.	FDGID
3454.	DGIDA
3455.	GIDAA
3456.	IDAAF
3457.	DAAFL
3458.	AAFLI
3459.	AFLIG
3460.	FLIGS
3461.	LIGSV
3462.	IGSVP
3463.	GSVPR
3464.	SVPRG
3465.	VPRGP
3466.	PRGPG
3467.	RGPGM
3468.	GPGME
3469.	PGMER
3470.	GMERR
3471.	MERRD
3472.	ERRDL
3473.	RRDLL
3474.	RDLLK
3475.	DLLKK
3476.	LLKKN
3477.	LKKNG
3478.	KKNGE
3479.	KNGEI
3480.	NGEIF
3481.	GEIFA
3482.	EIFAT
3483.	IFATQ
3484.	FATQG
3485.	ATQGK
3486.	TQGKA
3487.	QGKAL
3488.	GKALN
3489.	KALNT
3490.	ALNTT
3491.	LNTTA
3492.	NTTAK
3493.	TTAKR
3494.	TAKRD
3495.	AKRDA
3496.	KRDAK
3497.	RDAKI
3498.	DAKIF
3499.	AKIFV
3500.	KIFVV
3501.	IFVVG
3502.	FVVGN
3503.	VVGNP
3504.	VGNPV
3505.	GNPVN
3506.	NPVNT
3507.	PVNTN
3508.	VNTNC
3509.	NTNCW
3510.	TNCWI
3511.	NCWIA
3512.	CWIAM
3513.	WIAMN
3514.	IAMNH
3515.	AMNHA
3516.	MNHAP
3517.	NHAPR
3518.	HAPRL
3519.	APRLL
3520.	PRLLR
3521.	RLLRK
3522.	LLRKN
3523.	LRKNF
3524.	RKNFH
3525.	KNFHA
3526.	NFHAM
3527.	FHAML
3528.	HAMLR
3529.	AMLRL
3530.	MLRLD
3531.	LRLDQ
3532.	RLDQN
3533.	LDQNR
3534.	DQNRM
3535.	QNRMH
3536.	NRMHS
3537.	RMHSM
3538.	MHSML
3539.	HSMLS
3540.	SMLSH
3541.	MLSHR
3542.	LSHRA
3543.	SHRAE
3544.	HRAEV
3545.	RAEVP
3546.	AEVPL
3547.	EVPLS
3548.	VPLSA
3549.	PLSAV
3550.	LSAVS
3551.	SAVSQ
3552.	AVSQV
3553.	VSQVV
3554.	SQVVV
3555.	QVVVW
3556.	VVVWG
3557.	VVWGN
3558.	VWGNH
3559.	WGNHS
3560.	GNHSA
3561.	NHSAK
3562.	HSAKQ
3563.	SAKQV
3564.	AKQVP
3565.	KQVPD
3566.	QVPDF
3567.	VPDFT
3568.	PDFTQ
3569.	DFTQA
3570.	FTQAL
3571.	TQALI
3572.	QALIN
3573.	ALIND
3574.	LINDR
3575.	INDRP
3576.	NDRPI
3577.	DRPIA
3578.	RPIAE
3579.	PIAET
3580.	IAETI
3581.	AETIA
3582.	ETIAD
3583.	TIADR
3584.	IADRD
3585.	ADRDW
3586.	DRDWL
3587.	RDWLE
3588.	DWLEN
3589.	WLENI
3590.	LENIM
3591.	ENIMV
3592.	NIMVP
3593.	IMVPS
3594.	MVPSV
3595.	VPSVQ
3596.	PSVQS
3597.	SVQSR
3598.	VQSRG
3599.	QSRGS
3600.	SRGSA
3601.	RGSAV
3602.	GSAVI
3603.	SAVIE
3604.	AVIEA
3605.	VIEAR
3606.	IEARG
3607.	EARGK
3608.	ARGKS
3609.	RGKSS
3610.	GKSSA
3611.	KSSAA
3612.	SSAAS
3613.	SAASA
3614.	AASAA
3615.	ASAAR
3616.	SAARA
3617.	AARAL
3618.	ARALA
3619.	RALAE
3620.	ALAEA
3621.	LAEAA
3622.	AEAAR
3623.	EAARS
3624.	AARSI
3625.	ARSIY
3626.	RSIYQ
3627.	SIYQP
3628.	IYQPK
3629.	YQPKE
3630.	QPKEG
3631.	PKEGE
3632.	KEGEW
3633.	EGEWF
3634.	GEWFS
3635.	EWFSS
3636.	WFSSG
3637.	FSSGV
3638.	SSGVC
3639.	SGVCS
3640.	GVCSD
3641.	VCSDH
3642.	CSDHN
3643.	SDHNP
3644.	DHNPY
3645.	HNPYG
3646.	NPYGL
3647.	PYGLP
3648.	YGLPE
3649.	GLPED
3650.	LPEDL
3651.	PEDLI
3652.	EDLIF
3653.	DLIFG
3654.	LIFGF
3655.	IFGFP
3656.	FGFPC
3657.	GFPCR
3658.	FPCRM
3659.	PCRML
3660.	CRMLA
3661.	RMLAT
3662.	MLATG
3663.	LATGE
3664.	ATGEY
3665.	TGEYE
3666.	GEYEV
3667.	EYEVI
3668.	YEVIP
3669.	EVIPR
3670.	VIPRL
3671.	IPRLP
3672.	PRLPW
3673.	RLPWD
3674.	LPWDA
3675.	PWDAF
3676.	WDAFI
3677.	DAFIR
3678.	AFIRG
3679.	FIRGK
3680.	IRGKM
3681.	RGKMQ
3682.	GKMQI
3683.	KMQIS
3684.	MQISL
3685.	QISLD
3686.	ISLDE
3687.	SLDEI
3688.	LDEIL
3689.	DEILQ
3690.	EILQE
3691.	ILQEK
3692.	LQEKA
3693.	QEKAS
3694.	EKASV
3695.	KASVS
3696.	ASVSL

5-mer peptides of CT557 (dihydrolipoamide

dehydrogenase, 465 amino acids), ACCESSION# NP

220072.1, GI: 15605286 (SEQ ID NOS: 3697-4018)

3697.	MVSQT
3698.	VSQTV
3699.	SQTVS
3700.	QTVSV
3701.	TVSVA
3702.	VSVAV
3703.	SVAVT
3704.	VAVTG
3705.	AVTGG
3706.	VTGGT
3707.	TGGTG
3708.	GGTGQ
3709.	GTGQI
3710.	TGQIA
3711.	GQIAY
3712.	QIAYS
3713.	IAYSF
3714.	AYSFL
3715.	YSFLF
3716.	SFLFS
3717.	FLFSL
3718.	LFSLA
3719.	FSLAH
3720.	SLAHG
3721.	LAHGD
3722.	AHGDV
3723.	HGDVF
3724.	GDVFG
3725.	DVFGL
3726.	VFGLD
3727.	FGLDC
3728.	GLDCG
3729.	LDCGI
3730.	DCGID
3731.	CGIDL
3732.	GIDLR
3733.	IDLRI
3734.	DLRIY
3735.	LRIYD
3736.	RIYDI
3737.	IYDIP
3738.	YDIPG
3739.	DIPGT
3740.	IPGTE
3741.	PGTER
3742.	GTERA
3743.	TERAL
3744.	ERALS
3745.	RALSG
3746.	ALSGV
3747.	LSGVR
3748.	SGVRM
3749.	GVRME
3750.	VRMEL
3751.	RMELD
3752.	MELDD
3753.	ELDDG
3754.	LDDGA
3755.	DDGAF
3756.	DGAFP
3757.	GAFPL
3758.	AFPLL
3759.	FPLLQ
3760.	PLLQR
3761.	LLQRV
3762.	LQRVQ
3763.	QRVQV
3764.	RVQVT
3765.	VQVTT
3766.	QVTTS
3767.	VTTSL
3768.	TTSLH
3769.	TSLHD
3770.	SLHDA
3771.	LHDAF
3772.	HDAFD
3773.	DAFDG
3774.	AFDGI
3775.	FDGID
3776.	DGIDA
3777.	GIDAA
3778.	IDAAF
3779.	DAAFL
3780.	AAFLI
3781.	AFLIG
3782.	FLIGS
3783.	LIGSV
3784.	IGSVP
3785.	GSVPR
3786.	SVPRG
3787.	VPRGP
3788.	PRGPG
3789.	RGPGM
3790.	GPGME
3791.	PGMER
3792.	GMERR
3793.	MERRD
3794.	ERRDL
3795.	RRDLL
3796.	RDLLK
3797.	DLLKK
3798.	LLKKN
3799.	LKKNG
3800.	KKNGE
3801.	KNGEI
3802.	NGEIF
3803.	GEIFA
3804.	EIFAT
3805.	IFATQ
3806.	FATQG
3807.	ATQGK
3808.	TQGKA
3809.	QGKAL
3810.	GKALN
3811.	KALNT
3812.	ALNTT
3813.	LNTTA
3814.	NTTAK
3815.	TTAKR
3816.	TAKRD
3817.	AKRDA
3818.	KRDAK
3819.	RDAKI
3820.	DAKIF
3821.	AKIFV
3822.	KIFVV
3823.	IFVVG
3824.	FVVGN
3825.	VVGNP
3826.	VGNPV
3827.	GNPVN
3828.	NPVNT
3829.	PVNTN
3830.	VNTNC
3831.	NTNCW
3832.	TNCWI
3833.	NCWIA
3834.	CWIAM
3835.	WIAMN
3836.	IAMNH
3837.	AMNHA
3838.	MNHAP
3839.	NHAPR
3840.	HAPRL
3841.	APRLL
3842.	PRLLR
3843.	RLLRK
3844.	LLRKN
3845.	LRKNF
3846.	RKNFH
3847.	KNFHA
3848.	NFHAM
3849.	FHAML
3850.	HAMLR
3851.	AMLRL
3852.	MLRLD
3853.	LRLDQ
3854.	RLDQN
3855.	LDQNR
3856.	DQNRM
3857.	QNRMH
3858.	NRMHS
3859.	RMHSM
3860.	MHSML
3861.	HSMLS
3862.	SMLSH
3863.	MLSHR
3864.	LSHRA
3865.	SHRAE
3866.	HRAEV
3867.	RAEVP
3868.	AEVPL
3869.	EVPLS
3870.	VPLSA
3871.	PLSAV
3872.	LSAVS
3873.	SAVSQ
3874.	AVSQV
3875.	VSQVV
3876.	SQVVV
3877.	QVVVW
3878.	VVVWG
3879.	VVWGN
3880.	VWGNH
3881.	WGNHS
3882.	GNHSA
3883.	NHSAK
3884.	HSAKQ
3885.	SAKQV
3886.	AKQVP
3887.	KQVPD
3888.	QVPDF
3889.	VPDFT
3890.	PDFTQ
3891.	DFTQA
3892.	FTQAL
3893.	TQALI
3894.	QALIN
3895.	ALIND
3896.	LINDR
3897.	INDRP
3898.	NDRPI
3899.	DRPIA
3900.	RPIAE
3901.	PIAET
3902.	IAETI
3903.	AETIA
3904.	ETIAD
3905.	TIADR
3906.	IADRD
3907.	ADRDW
3908.	DRDWL
3909.	RDWLE
3910.	DWLEN
3911.	WLENI
3912.	LENIM
3913.	ENIMV
3914.	NIMVP
3915.	IMVPS
3916.	MVPSV
3917.	VPSVQ
3918.	PSVQS
3919.	SVQSR
3920.	VQSRG
3921.	QSRGS
3922.	SRGSA
3923.	RGSAV
3924.	GSAVI
3925.	SAVIE
3926.	AVIEA
3927.	VIEAR
3928.	IEARG
3929.	EARGK
3930.	ARGKS
3931.	RGKSS
3932.	GKSSA
3933.	KSSAA
3934.	SSAAS
3935.	SAASA
3936.	AASAA
3937.	ASAAR
3938.	SAARA
3939.	AARAL
3940.	ARALA
3941.	RALAE
3942.	ALAEA
3943.	LAEAA
3944.	AEAAR
3945.	EAARS
3946.	AARSI
3947.	ARSIY
3948.	RSIYQ
3949.	SIYQP
3950.	IYQPK
3951.	YQPKE
3952.	QPKEG
3953.	PKEGE
3954.	KEGEW
3955.	EGEWF
3956.	GEWFS
3957.	EWFSS
3958.	WFSSG
3959.	FSSGV
3960.	SSGVC
3961.	SGVCS
3962.	GVCSD
3963.	VCSDH
3964.	CSDHN
3965.	SDHNP
3966.	DHNPY
3967.	HNPYG
3968.	NPYGL
3969.	PYGLP
3970.	YGLPE
3971.	GLPED
3972.	LPEDL
3973.	PEDLI
3974.	EDLIF
3975.	DLIFG
3976.	LIEGE
3977.	IFGFP
3978.	FGFPC
3979.	GFPCR
3980.	FPCRM
3981.	PCRML
3982.	CRMLA
3983.	RMLAT
3984.	MLATG
3985.	LATGE
3986.	ATGEY
3987.	TGEYE
3988.	GEYEV
3989.	EYEVI
3990.	YEVIP
3991.	EVIPR
3992.	VIPRL
3993.	IPRLP
3994.	PRLPW
3995.	RLPWD
3996.	LPWDA
3997.	PWDAF
3998.	WDAFI
3999.	DAFIR
4000.	AFIRG
4001.	FIRGK
4002.	IRGKM
4003.	RGKMQ
4004.	GKMQI
4005.	KMQIS
4006.	MQISL
4007.	QISLD
4008.	ISLDE
4009.	SLDEI
4010.	LDEIL
4011.	DEILQ
4012.	EILQE
4013.	ILQEK
4014.	LQEKA
4015.	QEKAS
4016.	EKASV
4017.	KASVS
4018.	ASVSL

5-mer peptides of CT858 (CPAF, total of 609 amino

acids), ACCESSION# AAC68456.1, GI: 3329332

(SEQ ID NOS: 4019-4623)

4019.	MGFWR
4020.	GFWRT
4021.	FWRTS
4022.	WRTSI
4023.	RTSIM
4024.	TSIMK
4025.	SIMKM
4026.	IMKMN
4027.	MKMNR
4028.	KMNRI
4029.	MNRIW
4030.	NRIWL
4031.	RIWLL
4032.	IWLLL
4033.	WLLLL
4034.	LLLLT
4035.	LLLTF
4036.	LLTFS
4037.	LTFSS
4038.	TFSSA
4039.	FSSAI
4040.	SSAIH
4041.	SAIHS
4042.	AIHSP
4043.	IHSPV
4044.	HSPVQ
4045.	SPVQG
4046.	PVQGE
4047.	VQGES
4048.	QGESL
4049.	GESLV
4050.	ESLVC
4051.	SLVCK
4052.	LVCKN
4053.	VCKNA
4054.	CKNAL
4055.	KNALQ
4056.	NALQD
4057.	ALQDL
4058.	LQDLS
4059.	QDLSF
4060.	DLSFL
4061.	LSFLE
4062.	SFLEH
4063.	FLEHL
4064.	LEHLL
4065.	EHLLQ
4066.	HLLQV
4067.	LLQVK
4068.	LQVKY
4069.	QVKYA
4070.	VKYAP
4071.	KYAPK
4072.	YAPKT
4073.	APKTW
4074.	PKTWK
4075.	KTWKE
4076.	TWKEQ
4077.	WKEQY
4078.	KEQYL
4079.	EQYLG
4080.	QYLGW
4081.	YLGWD
4082.	LGWDL
4083.	GWDLV
4084.	WDLVQ
4085.	DLVQS
4086.	LVQSS
4087.	VQSSV
4088.	QSSVS
4089.	SSVSA
4090.	SVSAQ
4091.	VSAQQ
4092.	SAQQK
4093.	AQQKL
4094.	QQKLR
4095.	QKLRT
4096.	KLRTQ
4097.	LRTQE
4098.	RTQEN
4099.	TQENP
4100.	QENPS
4101.	ENPST
4102.	NPSTS
4103.	PSTSF
4104.	STSFC
4105.	TSFCQ
4106.	SFCQQ
4107.	FCQQV
4108.	CQQVL
4109.	QQVLA
4110.	QVLAD
4111.	VLADF
4112.	LADFI
4113.	ADFIG
4114.	DFIGG
4115.	FIGGL
4116.	IGGLN
4117.	GGLND
4118.	GLNDF
4119.	LNDFH
4120.	NDFHA
4121.	DFHAG
4122.	FHAGV
4123.	HAGVT
4124.	AGVTF
4125.	GVTFF
4126.	VTFFA
4127.	TFFAI
4128.	FFAIE
4129.	FATES
4130.	AIESA
4131.	IESAY
4132.	ESAYL
4133.	SAYLP
4134.	AYLPY
4135.	YLPYT
4136.	LPYTV
4137.	PYTVQ
4138.	YTVQK
4139.	TVQKS
4140.	VQKSS
4141.	QKSSD
4142.	KSSDG
4143.	SSDGR
4144.	SDGRF
4145.	DGRFY
4146.	GRFYF
4147.	RFYFV
4148.	FYFVD
4149.	YFVDI
4150.	FVDIM
4151.	VDIMT
4152.	DIMTF
4153.	IMTFS
4154.	MTFSS
4155.	TFSSE
4156.	FSSEI
4157.	SSEIR
4158.	SEIRV
4159.	EIRVG
4160.	IRVGD
4161.	RVGDE
4162.	VGDEL
4163.	GDELL
4164.	DELLE
4165.	ELLEV
4166.	LLEVD
4167.	LEVDG
4168.	EVDGA
4169.	VDGAP
4170.	DGAPV
4171.	GAPVQ
4172.	APVQD
4173.	PVQDV
4174.	VQDVL
4175.	QDVLA
4176.	DVLAT
4177.	VLATL
4178.	LATLY
4179.	ATLYG
4180.	TLYGS
4181.	LYGSN
4182.	YGSNH
4183.	GSNHK
4184.	SNHKG
4185.	NHKGT
4186.	HKGTA
4187.	KGTAA
4188.	GTAAE
4189.	TAAEE
4190.	AAEES
4191.	AEESA
4192.	EESAA
4193.	ESAAL
4194.	SAALR
4195.	AALRT
4196.	ALRTL
4197.	LRTLF
4198.	RTLFS
4199.	TLFSR
4200.	LFSRM
4201.	FSRMA
4202.	SRMAS
4203.	RMASL
4204.	MASLG
4205.	ASLGH
4206.	SLGHK
4207.	LGHKV
4208.	GHKVP
4209.	HKVPS
4210.	KVPSG
4211.	VPSGR
4212.	PSGRT
4213.	SGRTT
4214.	GRTTL
4215.	RTTLK
4216.	TTLKI
4217.	TLKIR
4218.	LKIRR
4219.	KIRRP
4220.	IRRPF
4221.	RRPFG
4222.	RPFGT
4223.	PFGTT
4224.	FGTTR
4225.	GTTRE
4226.	TTREV
4227.	TREVR
4228.	REVRV
4229.	EVRVK
4230.	VRVKW
4231.	RVKWR
4232.	VKWRY
4233.	KWRYV
4234.	WRYVP
4235.	RYVPE
4236.	YVPEG
4237.	VPEGV
4238.	PEGVG
4239.	EGVGD
4240.	GVGDL
4241.	VGDLA
4242.	GDLAT
4243.	DLATI
4244.	LATIA
4245.	ATIAP
4246.	TIAPS
4247.	IAPSI
4248.	APSIR
4249.	PSIRA
4250.	STRAP
4251.	IRAPQ
4252.	RAPQL
4253.	APQLQ
4254.	PQLQK
4255.	QLQKS
4256.	LQKSM
4257.	QKSMR
4258.	KSMRS
4259.	SMRSF
4260.	MRSFF
4261.	RSFFP
4262.	SFFPK
4263.	FFPKK
4264.	FPKKD
4265.	PKKDD
4266.	KKDDA
4267.	KDDAF
4268.	DDAFH
4269.	DAFHR
4270.	AFHRS
4271.	FHRSS
4272.	HRSSS
4273.	RSSSL
4274.	SSSLF
4275.	SSLFY
4276.	SLFYS
4277.	LFYSP
4278.	FYSPM
4279.	YSPMV
4280.	SPMVP
4281.	PMVPH
4282.	MVPHF
4283.	VPHFW
4284.	PHFWA
4285.	HFWAE
4286.	FWAEL
4287.	WAELR
4288.	AELRN
4289.	ELRNH
4290.	LRNHY
4291.	RNHYA
4292.	NHYAT
4293.	HYATS
4294.	YATSG
4295.	ATSGL
4296.	TSGLK
4297.	SGLKS
4298.	GLKSG
4299.	LKSGY
4300.	KSGYN
4301.	SGYNI
4302.	GYNIG
4303.	YNIGS
4304.	NIGST
4305.	IGSTD
4306.	GSTDG
4307.	STDGF
4308.	TDGFL
4309.	DGFLP
4310.	GFLPV
4311.	FLPVI
4312.	LPVIG
4313.	PVIGP
4314.	VIGPV
4315.	IGPVI
4316.	GPVIW
4317.	PVIWE
4318.	VIWES
4319.	IWESE
4320.	WESEG
4321.	ESEGL
4322.	SEGLF
4323.	EGLFR
4324.	GLFRA
4325.	LFRAY
4326.	FRAYI
4327.	RAYIS
4328.	AYISS
4329.	YISSV
4330.	ISSVT
4331.	SSVTD
4332.	SVTDG
4333.	VTDGD
4334.	TDGDG
4335.	DGDGK
4336.	GDGKS
4337.	DGKSH
4338.	GKSHK
4339.	KSHKV
4340.	SHKVG
4341.	HKVGF
4342.	KVGFL
4343.	VGFLR
4344.	GFLRI
4345.	FLRIP
4346.	LRIPT
4347.	RIPTY
4348.	IPTYS
4349.	PTYSW
4350.	TYSWQ
4351.	YSWQD
4352.	SWQDM
4353.	WQDME
4354.	QDMED
4355.	DMEDF
4356.	MEDFD
4357.	EDFDP
4358.	DFDPS
4359.	FDPSG
4360.	DPSGP
4361.	PSGPP
4362.	SGPPP
4363.	GPPPW
4364.	PPPWE
4365.	PPWEE
4366.	PWEEF
4367.	WEEFA
4368.	EEFAK
4369.	EFAKI
4370.	FAKII
4371.	AKIIQ
4372.	KIIQV
4373.	IIQVF
4374.	IQVFS
4375.	QVFSS
4376.	VFSSN
4377.	FSSNT
4378.	SSNTE
4379.	SNTEA
4380.	NTEAL
4381.	TEALI
4382.	EALII
4383.	ALIID
4384.	LIIDQ
4385.	IIDQT
4386.	IDQTN
4387.	DQTNN
4388.	QTNNP
4389.	TNNPG
4390.	NNPGG
4391.	NPGGS
4392.	PGGSV
4393.	GGSVL
4394.	GSVLY
4395.	SVLYL
4396.	VLYLY
4397.	LYLYA
4398.	YLYAL
4399.	LYALL
4400.	YALLS
4401.	ALLSM
4402.	LLSML
4403.	LSMLT
4404.	SMLTD
4405.	MLTDR
4406.	LTDRP
4407.	TDRPL
4408.	DRPLE
4409.	RPLEL
4410.	PLELP
4411.	LELPK
4412.	ELPKH
4413.	LPKHR
4414.	PKHRM
4415.	KHRMI
4416.	HRMIL
4417.	RMILT
4418.	MILTQ
4419.	ILTQD
4420.	LTQDE
4421.	TQDEV
4422.	QDEVV
4423.	DEVVD
4424.	EVVDA
4425.	VVDAL
4426.	VDALD
4427.	DALDW
4428.	ALDWL
4429.	LDWLT
4430.	DWLTL
4431.	WLTLL
4432.	LTLLE
4433.	TLLEN
4434.	LLENV
4435.	LENVD
4436.	ENVDT
4437.	NVDTN
4438.	VDTNV
4439.	DTNVE
4440.	TNVES
4441.	NVESR
4442.	VESRL
4443.	ESRLA
4444.	SRLAL
4445.	RLALG
4446.	LALGD
4447.	ALGDN
4448.	LGDNM
4449.	GDNME
4450.	DNMEG
4451.	NMEGY
4452.	MEGYT
4453.	EGYTV
4454.	GYTVD
4455.	YTVDL
4456.	TVDLQ
4457.	VDLQV
4458.	DLQVA
4459.	LQVAE
4460.	QVAEY
4461.	VAEYL
4462.	AEYLK
4463.	EYLKS
4464.	YLKSF
4465.	LKSFG
4466.	KSFGR
4467.	SFGRQ
4468.	FGRQV
4469.	GRQVL
4470.	RQVLN
4471.	QVLNC
4472.	VLNCW
4473.	LNCWS
4474.	NCWSK
4475.	CWSKG
4476.	WSKGD
4477.	SKGDI
4478.	KGDIE
4479.	GDIEL
4480.	DIELS
4481.	IELST
4482.	ELSTP
4483.	LSTPI
4484.	STPIP
4485.	TPIPL
4486.	PIPLF
4487.	IPLFG
4488.	PLFGF
4489.	LFGFE
4490.	FGFEK
4491.	GFEKI
4492.	FEKIH
4493.	EKIHP
4494.	KIHPH
4495.	IHPHP
4496.	HPHPR
4497.	PHPRV
4498.	HPRVQ
4499.	PRVQY
4500.	RVQYS
4501.	VQYSK
4502.	QYSKP
4503.	YSKPI
4504.	SKPIC
4505.	KPICV
4506.	PICVL
4507.	ICVLI
4508.	CVLIN
4509.	VLINE
4510.	LINEQ
4511.	INEQD
4512.	NEQDF
4513.	EQDFS
4514.	QDFSC
4515.	DFSCA
4516.	FSCAD
4517.	SCADF
4518.	CADFF
4519.	ADFFP
4520.	DFFPV
4521.	FFPVV
4522.	FPVVL
4523.	PVVLK
4524.	VVLKD
4525.	VLKDN
4526.	LKDND
4527.	KDNDR
4528.	DNDRA
4529.	NDRAL
4530.	DRALI
4531.	RALIV
4532.	ALIVG
4533.	LIVGT
4534.	IVGTR
4535.	VGTRT
4536.	GTRTA
4537.	TRTAG
4538.	RTAGA
4539.	TAGAG
4540.	AGAGG
4541.	GAGGF
4542.	AGGFV
4543.	GGFVF
4544.	GFVFN
4545.	FVFNV
4546.	VFNVQ
4547.	FNVQF
4548.	NVQFP
4549.	VQFPN
4550.	QFPNR
4551.	FPNRT
4552.	PNRTG
4553.	NRTGI
4554.	RTGIK
4555.	TGIKT
4556.	GIKTC
4557.	IKTCS
4558.	KTCSL
4559.	TCSLT
4560.	CSLTG
4561.	SLTGS
4562.	LTGSL
4563.	TGSLA
4564.	GSLAV
4565.	SLAVR
4566.	LAVRE
4567.	AVREH
4568.	VREHG
4569.	REHGA
4570.	EHGAF
4571.	HGAFI
4572.	GAFIE
4573.	AFIEN
4574.	FIENI
4575.	IENIG
4576.	ENIGV
4577.	NIGVE
4578.	IGVEP
4579.	GVEPH
4580.	VEPHI
4581.	EPHID
4582.	PHIDL
4583.	HIDLP
4584.	IDLPF
4585.	DLPFT
4586.	LPFTA
4587.	PFTAN
4588.	FTAND
4589.	TANDI
4590.	ANDIR
4591.	NDIRY
4592.	DIRYK
4593.	IRYKG
4594.	RYKGY
4595.	YKGYS
4596.	KGYSE
4597.	GYSEY
4598.	YSEYL
4599.	SEYLD
4600.	EYLDK
4601.	YLDKV
4602.	LDKVK
4603.	DKVKK
4604.	KVKKL
4605.	VKKLV
4606.	KKLVC
4607.	KLVCQ
4608.	LVCQL
4609.	VCQLI
4610.	CQLIN
4611.	QLINN
4612.	LINND
4613.	INNDG
4614.	NNDGT
4615.	NDGTI
4616.	DGTII
4617.	GTIIL
4618.	TIILA
4619.	IILAE
4620.	ILAED
4621.	LAEDG
4622.	AEDGS
4623.	EDGSF

5-mer peptides of Pgp3 (plasmid-encoded secretion

protein, total of 264 amino acids), ACCESSION#

ADI51551.1, GI: 297749006 (SEQ ID NOS: 4624-4883)

4624.	MGNSG
4625.	GNSGF
4626.	NSGFY
4627.	SGFYL
4628.	GFYLY
4629.	FYLYN
4630.	YLYNT
4631.	LYNTE
4632.	YNTEN
4633.	NTENC
4634.	TENCV
4635.	ENCVF
4636.	NCVFA
4637.	CVFAD
4638.	VFADN
4639.	FADNI
4640.	ADNIK
4641.	DNIKV
4642.	NIKVG
4643.	IKVGQ
4644.	KVGQM
4645.	VGQMT
4646.	GQMTE
4647.	QMTEP
4648.	MTEPL
4649.	TEPLK
4650.	EPLKD
4651.	PLKDQ
4652.	LKDQQ
4653.	KDQQI
4654.	DQQII
4655.	QQIIL
4656.	QIILG
4657.	IILGT
4658.	ILGTK
4659.	LGTKS
4660.	GTKST
4661.	TKSTP
4662.	KSTPV
4663.	STPVA
4664.	TPVAA
4665.	PVAAK
4666.	VAAKM
4667.	AAKMT
4668.	AKMTA
4669.	KMTAS
4670.	MTASD
4671.	TASDG
4672.	ASDGI
4673.	SDGIS
4674.	DGISL
4675.	GISLT
4676.	ISLTV
4677.	SLTVS
4678.	LTVSN
4679.	TVSNN
4680.	VSNNS
4681.	SNNSS
4682.	NNSST
4683.	NSSTN
4684.	SSTNA
4685.	STNAS
4686.	TNASI
4687.	NASIT
4688.	ASITI
4689.	SITIG
4690.	ITIGL
4691.	TIGLD
4692.	IGLDA
4693.	GLDAE
4694.	LDAEK
4695.	DAEKA
4696.	AEKAY
4697.	EKAYQ
4698.	KAYQL
4699.	AYQLI
4700.	YQLIL
4701.	QLILE
4702.	LILEK
4703.	ILEKL
4704.	LEKLG
4705.	EKLGN
4706.	KLGNQ
4707.	LGNQI
4708.	GNQIL
4709.	NQILD
4710.	QILDG
4711.	ILDGI
4712.	LDGIA
4713.	DGIAD
4714.	GIADT
4715.	IADTI
4716.	ADTIV
4717.	DTIVD
4718.	TIVDS
4719.	IVDST
4720.	VDSTV
4721.	DSTVQ
4722.	STVQD
4723.	TVQDI
4724.	VQDIL
4725.	QDILD
4726.	DILDK
4727.	ILDKI
4728.	LDKIT
4729.	DKITT
4730.	KITTD
4731.	ITTDP
4732.	TTDPS
4733.	TDPSL
4734.	DPSLG
4735.	PSLGL
4736.	SLGLL
4737.	LGLLK
4738.	GLLKA
4739.	LLKAF
4740.	LKAFN
4741.	KAFNN
4742.	AFNNF
4743.	FNNFP
4744.	NNFPI
4745.	NFPIT
4746.	FPITN
4747.	PITNK
4748.	ITNKI
4749.	TNKIQ
4750.	NKIQC
4751.	KIQCN
4752.	IQCNG
4753.	QCNGL
4754.	CNGLF
4755.	NGLFT
4756.	GLFTP
4757.	LFTPS
4758.	FTPSN
4759.	TPSNI
4760.	PSNIE
4761.	SNIET
4762.	NIETL
4763.	IETLL
4764.	ETLLG
4765.	TLLGG
4766.	LLGGT
4767.	LGGTE
4768.	GGTEI
4769.	GTEIG
4770.	TEIGK
4771.	EIGKF
4772.	IGKFT
4773.	GKFTV
4774.	KFTVT
4775.	FTVTP
4776.	TVTPK
4777.	VTPKS
4778.	TPKSS
4779.	PKSSG
4780.	KSSGS
4781.	SSGSM
4782.	SGSMF
4783.	GSMFL
4784.	SMFLV
4785.	MFLVS
4786.	FLVSA
4787.	LVSAD
4788.	VSADI
4789.	SADII
4790.	ADIIA
4791.	DIIAS
4792.	IIASR
4793.	IASRM
4794.	ASRME
4795.	SRMEG
4796.	RMEGG
4797.	MEGGV
4798.	EGGVV
4799.	GGVVL
4800.	GVVLA
4801.	VVLAL
4802.	VLALV
4803.	LALVR
4804.	ALVRE
4805.	LVREG
4806.	VREGD
4807.	REGDS
4808.	EGDSK
4809.	GDSKP
4810.	DSKPC
4811.	SKPCA
4812.	KPCAI
4813.	PCAIS
4814.	CAISY
4815.	AISYG
4816.	ISYGY
4817.	SYGYS
4818.	YGYSS
4819.	GYSSG
4820.	YSSGV
4821.	SSGVP
4822.	SGVPN
4823.	GVPNL
4824.	VPNLC
4825.	PNLCS
4826.	NLCSL
4827.	LCSLR
4828.	CSLRT
4829.	SLRTS
4830.	LRTSI
4831.	RTSIT
4832.	TSITN
4833.	SITNT
4834.	ITNTG
4835.	TNTGL
4836.	NTGLT
4837.	TGLTP
4838.	GLTPT
4839.	LTPTT
4840.	TPTTY
4841.	PTTYS
4842.	TTYSL
4843.	TYSLR
4844.	YSLRV
4845.	SLRVG
4846.	LRVGG
4847.	RVGGL
4848.	VGGLE
4849.	GGLES
4850.	GLESG
4851.	LESGV
4852.	ESGVV
4853.	SGVVW
4854.	GVVWV
4855.	VVWVN
4856.	VWVNA
4857.	WVNAL
4858.	VNALS
4859.	NALSN
4860.	ALSNG
4861.	LSNGN
4862.	SNGND
4863.	NGNDI
4864.	GNDIL
4865.	NDILG
4866.	DILGI
4867.	ILGIT
4868.	LGITN
4869.	GITNT
4870.	ITNTS
4871.	TNTSN
4872.	NTSNV
4873.	TSNVS
4874.	SNVSF
4875.	NVSFL
4876.	VSFLE
4877.	SFLEV
4878.	FLEVI
4879.	LEVIP
4880.	EVIPQ
4881.	VIPQT
4882.	IPQTN
4883.	PQTNA

5-mer peptides of CT823 (cHtrA, 497 amino acids),

ACCESSION# NP 220344.1, GI: 15605558 (SEQ ID NOS:

4884-5376)

4884.	MMKRL
4885.	MKRLL
4886.	KRLLC
4887.	RLLCV
4888.	LLCVL
4889.	LCVLL
4890.	CVLLS
4891.	VLLST
4892.	LLSTS
4893.	LSTSV
4894.	STSVF
4895.	TSVFS
4896.	SVFSS
4897.	VFSSP
4898.	FSSPM
4899.	SSPML
4900.	SPMLG
4901.	PMLGY
4902.	MLGYS
4903.	LGYSA
4904.	GYSAS
4905.	YSASK
4906.	SASKK
4907.	ASKKD
4908.	SKKDS
4909.	KKDSK
4910.	KDSKA
4911.	DSKAD
4912.	SKADI
4913.	KADIC
4914.	ADICL
4915.	DICLA
4916.	ICLAV
4917.	CLAVS
4918.	LAVSS
4919.	AVSSG
4920.	VSSGD
4921.	SSGDQ
4922.	SGDQE
4923.	GDQEV
4924.	DQEVS
4925.	QEVSQ
4926.	EVSQE
4927.	VSQED
4928.	SQEDL
4929.	QEDLL
4930.	EDLLK
4931.	DLLKE
4932.	LLKEV
4933.	LKEVS
4934.	KEVSR
4935.	EVSRG
4936.	VSRGF
4937.	SRGFS
4938.	RGFSR
4939.	GFSRV
4940.	FSRVA
4941.	SRVAA
4942.	RVAAK
4943.	VAAKA
4944.	AAKAT
4945.	AKATP
4946.	KATPG
4947.	ATPGV
4948.	TPGVV
4949.	PGVVY
4950.	GVVYI
4951.	VVYIE
4952.	VYIEN
4953.	YIENF
4954.	IENFP
4955.	ENFPK
4956.	NFPKT
4957.	FPKTG
4958.	PKTGN
4959.	KTGNQ
4960.	TGNQA
4961.	GNQAI
4962.	NQAIA
4963.	QAIAS
4964.	AIASP
4965.	IASPG
4966.	ASPGN
4967.	SPGNK
4968.	PGNKR
4969.	GNKRG
4970.	NKRGF
4971.	KRGFQ
4972.	RGFQE
4973.	GFQEN
4974.	FQENP
4975.	QENPF
4976.	ENPFD
4977.	NPFDY
4978.	PFDYF
4979.	FDYFN
4980.	DYFND
4981.	YFNDE
4982.	FNDEF
4983.	NDEFF
4984.	DEFFN
4985.	EFFNR
4986.	FFNRF
4987.	FNRFF
4988.	NRFFG
4989.	RFFGL
4990.	FFGLP
4991.	FGLPS
4992.	GLPSH
4993.	LPSHR
4994.	PSHRE
4995.	SHREQ
4996.	HREQQ
4997.	REQQR
4998.	EQQRP
4999.	QQRPQ
5000.	QRPQQ
5001.	RPQQR
5002.	PQQRD
5003.	QQRDA
5004.	QRDAV
5005.	RDAVR
5006.	DAVRG
5007.	AVRGT
5008.	VRGTG
5009.	RGTGF
5010.	GTGFI
5011.	TGFIV
5012.	GFIVS
5013.	FIVSE
5014.	IVSED
5015.	VSEDG
5016.	SEDGY
5017.	EDGYV
5018.	DGYVV
5019.	GYVVT
5020.	YVVTN
5021.	VVTNH
5022.	VTNHH
5023.	TNHHV
5024.	NHHVV
5025.	HHVVE
5026.	HVVED
5027.	VVEDA
5028.	VEDAG
5029.	EDAGK
5030.	DAGKI
5031.	AGKIH
5032.	GKIHV
5033.	KIHVT
5034.	IHVTL
5035.	HVTLH
5036.	VTLHD
5037.	TLHDG
5038.	LHDGQ
5039.	HDGQK
5040.	DGQKY
5041.	GQKYT
5042.	QKYTA
5043.	KYTAK
5044.	YTAKI
5045.	TAKIV
5046.	AKIVG
5047.	KIVGL
5048.	IVGLD
5049.	VGLDP
5050.	GLDPK
5051.	LDPKT
5052.	DPKTD
5053.	PKTDL
5054.	KTDLA
5055.	TDLAV
5056.	DLAVI
5057.	LAVIK
5058.	AVIKI
5059.	VIKIQ
5060.	IKIQA
5061.	KIQAE
5062.	IQAEK
5063.	QAEKL
5064.	AEKLP
5065.	EKLPF
5066.	KLPFL
5067.	LPFLT
5068.	PFLTF
5069.	FLTFG
5070.	LTFGN
5071.	TFGNS
5072.	FGNSD
5073.	GNSDQ
5074.	NSDQL
5075.	SDQLQ
5076.	DQLQI
5077.	QLQIG
5078.	LQIGD
5079.	QIGDW
5080.	IGDWA
5081.	GDWAI
5082.	DWAIA
5083.	WAIAI
5084.	AIAIG
5085.	IAIGN
5086.	AIGNP
5087.	IGNPF
5088.	GNPFG
5089.	NPFGL
5090.	PFGLQ
5091.	FGLQA
5092.	GLQAT
5093.	LQATV
5094.	QATVT
5095.	ATVTV
5096.	TVTVG
5097.	VTVGV
5098.	TVGVI
5099.	VGVIS
5100.	GVISA
5101.	VISAK
5102.	ISAKG
5103.	SAKGR
5104.	AKGRN
5105.	KGRNQ
5106.	GRNQL
5107.	RNQLH
5108.	NQLHI
5109.	QLHIV
5110.	LHIVD
5111.	HIVDF
5112.	IVDFE
5113.	VDFED
5114.	DFEDF
5115.	FEDFI
5116.	EDFIQ
5117.	DFIQT
5118.	FIQTD
5119.	IQTDA
5120.	QTDAA
5121.	TDAAI
5122.	DAAIN
5123.	AAINP
5124.	AINPG
5125.	INPGN
5126.	NPGNS
5127.	PGNSG
5128.	GNSGG
5129.	NSGGP
5130.	SGGPL
5131.	GGPLL
5132.	GPLLN
5133.	PLLNI
5134.	LLNIN
5135.	LNING
5136.	NINGQ
5137.	INGQV
5138.	NGQVI
5139.	GQVIG
5140.	QVIGV
5141.	VIGVN
5142.	IGVNT
5143.	GVNTA
5144.	VNTAI
5145.	NTAIV
5146.	TAIVS
5147.	AIVSG
5148.	IVSGS
5149.	VSGSG
5150.	SGSGG
5151.	GSGGY
5152.	SGGYI
5153.	GGYIG
5154.	GYIGI
5155.	YIGIG
5156.	IGIGF
5157.	GIGFA
5158.	IGFAI
5159.	GFAIP
5160.	FAIPS
5161.	AIPSL
5162.	IPSLM
5163.	PSLMA
5164.	SLMAK
5165.	LMAKR
5166.	MAKRV
5167.	AKRVI
5168.	KRVID
5169.	RVIDQ
5170.	VIDQL
5171.	IDQLI
5172.	DQLIS
5173.	QLISD
5174.	LISDG
5175.	ISDGQ
5176.	SDGQV
5177.	DGQVT
5178.	GQVTR
5179.	QVTRG
5180.	VTRGF
5181.	TRGFL
5182.	RGFLG
5183.	GFLGV
5184.	FLGVT
5185.	LGVTL
5186.	GVTLQ
5187.	VTLQP
5188.	TLQPI
5189.	LQPID
5190.	QPIDS
5191.	PIDSE
5192.	IDSEL
5193.	DSELA
5194.	SELAT
5195.	ELATC
5196.	LATCY
5197.	ATCYK
5198.	TCYKL
5199.	CYKLE
5200.	YKLEK
5201.	KLEKV
5202.	LEKVY
5203.	EKVYG
5204.	KVYGA
5205.	VYGAL
5206.	YGALV
5207.	GALVT
5208.	ALVTD
5209.	LVTDV
5210.	VTDVV
5211.	TDVVK
5212.	DVVKG
5213.	VVKGS
5214.	VKGSP
5215.	KGSPA
5216.	GSPAE
5217.	SPAEK
5218.	PAEKA
5219.	AEKAG
5220.	EKAGL
5221.	KAGLR
5222.	AGLRQ
5223.	GLRQE
5224.	LRQED
5225.	RQEDV
5226.	QEDVI
5227.	EDVIV
5228.	DVIVA
5229.	VIVAY
5230.	IVAYN
5231.	VAYNG
5232.	AYNGK
5233.	YNGKE
5234.	NGKEV
5235.	GKEVE
5236.	KEVES
5237.	EVESL
5238.	VESLS
5239.	ESLSA
5240.	SLSAL
5241.	LSALR
5242.	SALRN
5243.	ALRNA
5244.	LRNAI
5245.	RNAIS
5246.	NAISL
5247.	AISLM
5248.	ISLMM
5249.	SLMMP
5250.	LMMPG
5251.	MMPGT
5252.	MPGTR
5253.	PGTRV
5254.	GTRVV
5255.	TRVVL
5256.	RVVLK
5257.	VVLKI
5258.	VLKIV
5259.	LKIVR
5260.	KIVRE
5261.	IVREG
5262.	VREGK
5263.	REGKT
5264.	EGKTI
5265.	GKTIE
5266.	KTIEI
5267.	TIEIP
5268.	IEIPV
5269.	EIPVT
5270.	IPVTV
5271.	PVTVT
5272.	VTVTQ
5273.	TVTQI
5274.	VTQIP
5275.	TQIPT
5276.	QIPTE
5277.	IPTED
5278.	PTEDG
5279.	TEDGV
5280.	EDGVS
5281.	DGVSA
5282.	GVSAL
5283.	VSALQ
5284.	SALQK
5285.	ALQKM
5286.	LQKMG
5287.	QKMGV
5288.	KMGVR
5289.	MGVRV
5290.	GVRVQ
5291.	VRVQN
5292.	RVQNI
5293.	VQNIT
5294.	QNITP
5295.	NITPE
5296.	ITPEI
5297.	TPEIC
5298.	PEICK
5299.	EICKK
5300.	ICKKL
5301.	CKKLG
5302.	KKLGL
5303.	KLGLA
5304.	LGLAA
5305.	GLAAD
5306.	LAADT
5307.	AADTR
5308.	ADTRG
5309.	DTRGI
5310.	TRGIL
5311.	RGILV
5312.	GILVV
5313.	ILVVA
5314.	LVVAV
5315.	VVAVE
5316.	VAVEA
5317.	AVEAG
5318.	VEAGS
5319.	EAGSP
5320.	AGSPA
5321.	GSPAA
5322.	SPAAS
5323.	PAASA
5324.	AASAG
5325.	ASAGV
5326.	SAGVA
5327.	AGVAP
5328.	GVAPG
5329.	VAPGQ
5330.	APGQL
5331.	PGQLI
5332.	GQLIL
5333.	QLILA
5334.	LILAV
5335.	ILAVN
5336.	LAVNR
5337.	AVNRQ
5338.	VNRQR
5339.	NRQRV
5340.	RQRVA
5341.	QRVAS
5342.	RVASV
5343.	VASVE
5344.	ASVEE
5345.	SVEEL
5346.	VEELN
5347.	EELNQ
5348.	ELNQV
5349.	LNQVL
5350.	NQVLK
5351.	QVLKN
5352.	VLKNS
5353.	LKNSK
5354.	KNSKG
5355.	NSKGE
5356.	SKGEN
5357.	KGENV
5358.	GENVL
5359.	ENVLL
5360.	NVLLM
5361.	VLLMV
5362.	LLMVS
5363.	LMVSQ
5364.	MVSQG
5365.	VSQGD
5366.	SQGDV
5367.	QGDVV
5368.	GDVVR
5369.	DVVRF
5370.	VVRFI
5371.	VRFIV
5372.	RFIVL
5373.	FIVLK
5374.	IVLKS
5375.	VLKSD
5376.	LKSDE

5-mer peptides of CT681 (MOMP, 393 amino acids),

ACCESSION# NP 220200.1, GI: 15605414 (SEQ ID NOS:

5377-5765)

5377.	MKKLL
5378.	KKLLK
5379.	KLLKS
5380.	LLKSV
5381.	LKSVL
5382.	KSVLV
5383.	SVLVF
5384.	VLVFA
5385.	LVFAA
5386.	VFAAL
5387.	FAALS
5388.	AALSS
5389.	ALSSA
5390.	LSSAS
5391.	SSASS
5392.	SASSL
5393.	ASSLQ
5394.	SSLQA
5395.	SLQAL
5396.	LQALP
5397.	QALPV
5398.	ALPVG
5399.	LPVGN
5400.	PVGNP
5401.	VGNPA
5402.	GNPAE
5403.	NPAEP
5404.	PAEPS
5405.	AEPSL
5406.	EPSLM
5407.	PSLMI
5408.	SLMID
5409.	LMIDG
5410.	MIDGI
5411.	IDGIL
5412.	DGILW
5413.	GILWE
5414.	ILWEG
5415.	LWEGF
5416.	WEGFG
5417.	EGFGG
5418.	GFGGD
5419.	FGGDP
5420.	GGDPC
5421.	GDPCD
5422.	DPCDP
5423.	PCDPC
5424.	CDPCA
5425.	DPCAT
5426.	PCATW
5427.	CATWC
5428.	ATWCD
5429.	TWCDA
5430.	WCDAI
5431.	CDAIS
5432.	DAISM
5433.	AISMR
5434.	ISMRV
5435.	SMRVG
5436.	MRVGY
5437.	RVGYY
5438.	VGYYG
5439.	GYYGD
5440.	YYGDF
5441.	YGDFV
5442.	GDFVF
5443.	DFVFD
5444.	FVFDR
5445.	VFDRV
5446.	FDRVL
5447.	DRVLK
5448.	RVLKT
5449.	VLKTD
5450.	LKTDV
5451.	KTDVN
5452.	TDVNK
5453.	DVNKE
5454.	VNKEF
5455.	NKEFQ
5456.	KEFQM
5457.	EFQMG
5458.	FQMGA
5459.	QMGAK
5460.	MGAKP
5461.	GAKPT
5462.	AKPTT
5463.	KPTTD
5464.	PTTDT
5465.	TTDTG
5466.	TDTGN
5467.	DTGNS
5468.	TGNSA
5469.	GNSAA
5470.	NSAAP
5471.	SAAPS
5472.	AAPST
5473.	APSTL
5474.	PSTLT
5475.	STLTA
5476.	TLTAR
5477.	LTARE
5478.	TAREN
5479.	ARENP
5480.	RENPA
5481.	ENPAY
5482.	NPAYG
5483.	PAYGR
5484.	AYGRH
5485.	YGRHM
5486.	GRHMQ
5487.	RHMQD
5488.	HMQDA
5489.	MQDAE
5490.	QDAEM
5491.	DAEMF
5492.	AEMFT
5493.	EMFTN
5494.	MFTNA
5495.	FTNAA
5496.	TNAAC
5497.	NAACM
5498.	AACMA
5499.	ACMAL
5500.	CMALN
5501.	MALNI
5502.	ALNIW
5503.	LNIWD
5504.	NIWDR
5505.	IWDRF
5506.	WDRFD
5507.	DRFDV
5508.	RFDVF
5509.	FDVFC
5510.	DVFCT
5511.	VFCTL
5512.	FCTLG
5513.	CTLGA
5514.	TLGAT
5515.	LGATS
5516.	GATSG
5517.	ATSGY
5518.	TSGYL
5519.	SGYLK
5520.	GYLKG
5521.	YLKGN
5522.	LKGNS
5523.	KGNSA
5524.	GNSAS
5525.	NSASF
5526.	SASFN
5527.	ASFNL
5528.	SFNLV
5529.	FNLVG
5530.	NLVGL
5531.	LVGLF
5532.	VGLFG
5533.	GLFGD
5534.	LFGDN
5535.	FGDNE
5536.	GDNEN
5537.	DNENQ
5538.	NENQK
5539.	ENQKT
5540.	NQKTV
5541.	QKTVK
5542.	KTVKA
5543.	TVKAE
5544.	VKAES
5545.	KAESV
5546.	AESVP
5547.	ESVPN
5548.	SVPNM
5549.	VPNMS
5550.	PNMSF
5551.	NMSFD
5552.	MSFDQ
5553.	SFDQS
5554.	FDQSV
5555.	DQSVV
5556.	QSVVE
5557.	SVVEL
5558.	VVELY
5559.	VELYT
5560.	ELYTD
5561.	LYTDT
5562.	YTDTT
5563.	TDTTF
5564.	DTTFA
5565.	TTFAW
5566.	TFAWS
5567.	FAWSV
5568.	AWSVG
5569.	WSVGA
5570.	SVGAR
5571.	VGARA
5572.	GARAA
5573.	ARAAL
5574.	RAALW
5575.	AALWE
5576.	ALWEC
5577.	LWECG
5578.	WECGC
5579.	ECGCA
5580.	CGCAT
5581.	GCATL
5582.	CATLG
5583.	ATLGA
5584.	TLGAS
5585.	LGASF
5586.	GASFQ
5587.	ASFQY
5588.	SFQYA
5589.	FQYAQ
5590.	QYAQS
5591.	YAQSK
5592.	AQSKP
5593.	QSKPK
5594.	SKPKV
5595.	KPKVE
5596.	PKVEE
5597.	KVEEL
5598.	VEELN
5599.	EELNV
5600.	ELNVL
5601.	LNVLC
5602.	NVLCN
5603.	VLCNA
5604.	LCNAA
5605.	CNAAE
5606.	NAAEF
5607.	AAEFT
5608.	AEFTI
5609.	EFTIN
5610.	FTINK
5611.	TINKP
5612.	INKPK
5613.	NKPKG
5614.	KPKGY
5615.	PKGYV
5616.	KGYVG
5617.	GYVGK
5618.	YVGKE
5619.	VGKEF
5620.	GKEFP
5621.	KEFPL
5622.	EFPLD
5623.	FPLDL
5624.	PLDLT
5625.	LDLTA
5626.	DLTAG
5627.	LTAGT
5628.	TAGTD
5629.	AGTDA
5630.	GTDAA
5631.	TDAAT
5632.	DAATG
5633.	AATGT
5634.	ATGTK
5635.	TGTKD
5636.	GTKDA
5637.	TKDAS
5638.	KDASI
5639.	DASID
5640.	ASIDY
5641.	SIDYH
5642.	IDYHE
5643.	DYHEW
5644.	YHEWQ
5645.	HEWQA
5646.	EWQAS
5647.	WQASL
5648.	QASLA
5649.	ASLAL
5650.	SLALS
5651.	LALSY
5652.	ALSYR
5653.	LSYRL
5654.	SYRLN
5655.	YRLNM
5656.	RLNMF
5657.	LNMFT
5658.	NMFTP
5659.	MFTPY
5660.	FTPYI
5661.	TPYIG
5662.	PYIGV
5663.	YIGVK
5664.	IGVKW
5665.	GVKWS
5666.	VKWSR
5667.	KWSRA
5668.	WSRAS
5669.	SRASF
5670.	RASFD
5671.	ASFDA
5672.	SFDAD
5673.	FDADT
5674.	DADTI
5675.	ADTIR
5676.	DTIRI
5677.	TIRIA
5678.	IRIAQ
5679.	RIAQP
5680.	IAQPK
5681.	AQPKS
5682.	QPKSA
5683.	PKSAT
5684.	KSATA
5685.	SATAI
5686.	ATAIF
5687.	TAIFD
5688.	AIFDT
5689.	IFDTT
5690.	FDTTT
5691.	DTTTL
5692.	TTTLN
5693.	TTLNP
5694.	TLNPT
5695.	LNPTI
5696.	NPTIA
5697.	PTIAG
5698.	TIAGA
5699.	IAGAG
5700.	AGAGD
5701.	GAGDV
5702.	AGDVK
5703.	GDVKT
5704.	DVKTG
5705.	VKTGA
5706.	KTGAE
5707.	TGAEG
5708.	GAEGQ
5709.	AEGQL
5710.	EGQLG
5711.	GQLGD
5712.	QLGDT
5713.	LGDTM
5714.	GDTMQ
5715.	DTMQI
5716.	TMQIV
5717.	MQIVS
5718.	QIVSL
5719.	IVSLQ
5720.	VSLQL
5721.	SLQLN
5722.	LQLNK
5723.	QLNKM
5724.	LNKMK
5725.	NKMKS
5726.	KMKSR
5727.	MKSRK
5728.	KSRKS
5729.	SRKSC
5730.	RKSCG
5731.	KSCGI
5732.	SCGIA
5733.	CGIAV
5734.	GIAVG
5735.	IAVGT
5736.	AVGTT
5737.	VGTTI
5738.	GTTIV
5739.	TTIVD
5740.	TIVDA
5741.	IVDAD
5742.	VDADK
5743.	DADKY
5744.	ADKYA
5745.	DKYAV
5746.	KYAVT
5747.	YAVTV
5748.	AVTVE
5749.	VTVET
5750.	TVETR
5751.	VETRL
5752.	ETRLI
5753.	TRLID
5754.	RLIDE
5755.	LIDER
5756.	IDERA
5757.	DERAA
5758.	ERAAH
5759.	RAAHV
5760.	AAHVN
5761.	AHVNA
5762.	HVNAQ
5763.	VNAQF
5764.	NAQFR
5765.	AQFRF

5-mer peptides of CT119 (IncA, 273 amino acids),

ACCESSION# NP219622.1, GI: 15604838 (SEQ ID NOS:

5766-6034)

5766.	MTTPT
5767.	TTPTL
5768.	TPTLI
5769.	PTLIV
5770.	TLIVT
5771.	LIVTP
5772.	IVTPP
5773.	VTPPS
5774.	TPPSP
5775.	PPSPP
5776.	PSPPA
5777.	SPPAP
5778.	PPAPS
5779.	PAPSY
5780.	APSYS
5781.	PSYSA
5782.	SYSAN
5783.	YSANR
5784.	SANRV
5785.	ANRVP
5786.	NRVPQ
5787.	RVPQP
5788.	VPQPS
5789.	PQPSL
5790.	QPSLM
5791.	PSLMD
5792.	SLMDK
5793.	LMDKI
5794.	MDKIK
5795.	DKIKK
5796.	KIKKI
5797.	IKKIA
5798.	KKIAA
5799.	KIAAI
5800.	IAAIA
5801.	AAIAS
5802.	AIASL
5803.	IASLI
5804.	ASLIL
5805.	SLILI
5806.	LILIG
5807.	ILIGT
5808.	LIGTI
5809.	IGTIG
5810.	GTIGF
5811.	TIGFL
5812.	IGFLA
5813.	GFLAL
5814.	FLALL
5815.	LALLG
5816.	ALLGH
5817.	LLGHL
5818.	LGHLV
5819.	GHLVG
5820.	HLVGF
5821.	LVGFL
5822.	VGFLI
5823.	GFLIA
5824.	FLIAP
5825.	LIAPQ
5826.	IAPQI
5827.	APQIT
5828.	PQITI
5829.	QITIV
5830.	ITIVL
5831.	TIVLL
5832.	IVLLA
5833.	VLLAL
5834.	LLALF
5835.	LALFI
5836.	ALFII
5837.	LFIIS
5838.	FIISL
5839.	IISLA
5840.	ISLAG
5841.	SLAGN
5842.	LAGNA
5843.	AGNAL
5844.	GNALY
5845.	NALYL
5846.	ALYLQ
5847.	LYLQK
5848.	YLQKT
5849.	LQKTA
5850.	QKTAN
5851.	KTANL
5852.	TANLH
5853.	ANLHL
5854.	NLHLY
5855.	LHLYQ
5856.	HLYQD
5857.	LYQDL
5858.	YQDLQ
5859.	QDLQR
5860.	DLQRE
5861.	LQREV
5862.	QREVG
5863.	REVGS
5864.	EVGSL
5865.	VGSLK
5866.	GSLKE
5867.	SLKEI
5868.	LKEIN
5869.	KEINF
5870.	EINFM
5871.	INFML
5872.	NFMLS
5873.	FMLSV
5874.	MLSVL
5875.	LSVLQ
5876.	SVLQK
5877.	VLQKE
5878.	LQKEF
5879.	QKEFL
5880.	KEFLH
5881.	EFLHL
5882.	FLHLS
5883.	LHLSK
5884.	HLSKE
5885.	LSKEF
5886.	SKEFA
5887.	KEFAT
5888.	EFATT
5889.	FATTS
5890.	ATTSK
5891.	TTSKD
5892.	TSKDL
5893.	SKDLS
5894.	KDLSA
5895.	DLSAV
5896.	LSAVS
5897.	SAVSQ
5898.	AVSQD
5899.	VSQDF
5900.	SQDFY
5901.	QDFYS
5902.	DFYSC
5903.	FYSCL
5904.	YSCLQ
5905.	SCLQG
5906.	CLQGF
5907.	LQGFR
5908.	QGFRD
5909.	GFRDN
5910.	FRDNY
5911.	RDNYK
5912.	DNYKG
5913.	NYKGF
5914.	YKGFE
5915.	KGFES
5916.	GFESL
5917.	FESLL
5918.	ESLLD
5919.	SLLDE
5920.	LLDEY
5921.	LDEYK
5922.	DEYKN
5923.	EYKNS
5924.	YKNST
5925.	KNSTE
5926.	NSTEE
5927.	STEEM
5928.	TEEMR
5929.	EEMRK
5930.	EMRKL
5931.	MRKLF
5932.	RKLFS
5933.	KLFSQ
5934.	LFSQE
5935.	FSQEI
5936.	SQEII
5937.	QEIIA
5938.	EIIAD
5939.	IIADL
5940.	IADLK
5941.	ADLKG
5942.	DLKGS
5943.	LKGSV
5944.	KGSVA
5945.	GSVAS
5946.	SVASL
5947.	VASLR
5948.	ASLRE
5949.	SLREE
5950.	LREEI
5951.	REEIR
5952.	EEIRF
5953.	EIRFL
5954.	IRFLT
5955.	RFLTP
5956.	FLTPL
5957.	LTPLA
5958.	TPLAE
5959.	PLAEE
5960.	LAEEV
5961.	AEEVR
5962.	EEVRR
5963.	EVRRL
5964.	VRRLA
5965.	RRLAH
5966.	RLAHN
5967.	LAHNQ
5968.	AHNQQ
5969.	HNQQS
5970.	NQQSL
5971.	QQSLT
5972.	QSLTV
5973.	SLTVV
5974.	LTVVI
5975.	TVVIE
5976.	VVIEE
5977.	VIEEL
5978.	IEELK
5979.	EELKT
5980.	ELKTI
5981.	LKTIR
5982.	KTIRD
5983.	TIRDS
5984.	IRDSL
5985.	RDSLR
5986.	DSLRD
5987.	SLRDE
5988.	LRDEI
5989.	RDEIG
5990.	DEIGQ
5991.	EIGQL
5992.	IGQLS
5993.	GQLSQ
5994.	QLSQL
5995.	LSQLS
5996.	SQLSK
5997.	QLSKT
5998.	LSKTL
5999.	SKTLT
6000.	KTLTS
6001.	TLTSQ
6002.	LTSQI
6003.	TSQIA
6004.	SQIAL
6005.	QIALQ
6006.	IALQR
6007.	ALQRK
6008.	LQRKE
6009.	QRKES
6010.	RKESS
6011.	KESSD
6012.	ESSDL
6013.	SSDLC
6014.	SDLCS
6015.	DLCSQ
6016.	LCSQI
6017.	CSQIR
6018.	SQIRE
6019.	QIRET
6020.	IRETL
6021.	RETLS
6022.	ETLSS
6023.	TLSSP
6024.	LSSPR
6025.	SSPRK
6026.	SPRKS
6027.	PRKSA
6028.	RKSAS
6029.	KSASP
6030.	SASPS
6031.	ASPST
6032.	SPSTK
6033.	PSTKS
6034.	STKSS

5-mer peptides of CT813 (inclusion membrane

protein, 264 amino acids), ACCESSION# NP 220333.1,

GI: 15605547 (SEQ ID NOS: 6035-6294)

6035.	MTTLP
6036.	TTLPN
6037.	TLPNN
6038.	LPNNC
6039.	PNNCT
6040.	NNCTS
6041.	NCTSN
6042.	CTSNS
6043.	TSNSN
6044.	SNSNS
6045.	NSNSI
6046.	SNSIN
6047.	NSINT
6048.	SINTF
6049.	INTFT
6050.	NTFTK
6051.	TFTKD
6052.	FTKDI
6053.	TKDIE
6054.	KDIEM
6055.	DIEMA
6056.	IEMAK
6057.	EMAKQ
6058.	MAKQI
6059.	AKQIQ
6060.	KQIQG
6061.	QIQGS
6062.	IQGSR
6063.	QGSRK
6064.	GSRKD
6065.	SRKDP
6066.	RKDPL
6067.	KDPLA
6068.	DPLAK
6069.	PLAKT
6070.	LAKTS
6071.	AKTSW
6072.	KTSWI
6073.	TSWIA
6074.	SWIAG
6075.	WIAGL
6076.	IAGLI
6077.	AGLIC
6078.	GLICV
6079.	LICVV
6080.	ICVVA
6081.	CVVAG
6082.	VVAGV
6083.	VAGVL
6084.	AGVLG
6085.	GVLGL
6086.	VLGLL
6087.	LGLLA
6088.	GLLAI
6089.	LLAIG
6090.	LAIGI
6091.	AIGIG
6092.	IGIGG
6093.	GIGGC
6094.	IGGCS
6095.	GGCSM
6096.	GCSMA
6097.	CSMAS
6098.	SMASG
6099.	MASGL
6100.	ASGLG
6101.	SGLGL
6102.	GLGLI
6103.	LGLIG
6104.	GLIGA
6105.	LIGAI
6106.	IGAIV
6107.	GAIVA
6108.	AIVAA
6109.	IVAAV
6110.	VAAVI
6111.	AAVIV
6112.	AVIVA
6113.	VIVAV
6114.	IVAVG
6115.	VAVGL
6116.	AVGLC
6117.	VGLCC
6118.	GLCCL
6119.	LCCLV
6120.	CCLVS
6121.	CLVSA
6122.	LVSAL
6123.	VSALC
6124.	SALCL
6125.	ALCLQ
6126.	LCLQV
6127.	CLQVE
6128.	LQVEK
6129.	QVEKS
6130.	VEKSQ
6131.	EKSQW
6132.	KSQWW
6133.	SQWWQ
6134.	QWWQK
6135.	WWQKE
6136.	WQKEF
6137.	QKEFE
6138.	KEFES
6139.	EFESW
6140.	FESWI
6141.	ESWIE
6142.	SWIEQ
6143.	WIEQK
6144.	IEQKS
6145.	EQKSQ
6146.	QKSQF
6147.	KSQFR
6148.	SQFRI
6149.	QFRIV
6150.	FRIVM
6151.	RIVMA
6152.	IVMAD
6153.	VMADM
6154.	MADML
6155.	ADMLK
6156.	DMLKA
6157.	MLKAN
6158.	LKANR
6159.	KANRK
6160.	ANRKL
6161.	NRKLQ
6162.	RKLQS
6163.	KLQSE
6164.	LQSEV
6165.	QSEVE
6166.	SEVEF
6167.	EVEFL
6168.	VEFLS
6169.	EFLSK
6170.	FLSKG
6171.	LSKGW
6172.	SKGWS
6173.	KGWSD
6174.	GWSDD
6175.	WSDDT
6176.	SDDTA
6177.	DDTAV
6178.	DTAVH
6179.	TAVHK
6180.	AVHKE
6181.	VHKED
6182.	HKEDV
6183.	KEDVT
6184.	EDVTK
6185.	DVTKY
6186.	VTKYE
6187.	TKYEQ
6188.	KYEQV
6189.	YEQVV
6190.	EQVVE
6191.	QVVEE
6192.	VVEEY
6193.	VEEYA
6194.	EEYAE
6195.	EYAEK
6196.	YAEKI
6197.	AEKIM
6198.	EKIME
6199.	KIMEL
6200.	IMELY
6201.	MELYE
6202.	ELYEE
6203.	LYEET
6204.	YEETG
6205.	EETGV
6206.	ETGVL
6207.	TGVLT
6208.	GVLTI
6209.	VLTIE
6210.	LTIEK
6211.	TIEKI
6212.	IEKIN
6213.	EKINL
6214.	KINLQ
6215.	INLQK
6216.	NLQKE
6217.	LQKEK
6218.	QKEKK
6219.	KEKKA
6220.	EKKAW
6221.	KKAWL
6222.	KAWLE
6223.	AWLEE
6224.	WLEEK
6225.	LEEKA
6226.	EEKAE
6227.	EKAEM
6228.	KAEME
6229.	AEMEQ
6230.	EMEQK
6231.	MEQKL
6232.	EQKLT
6233.	QKLTT
6234.	KLTTV
6235.	LTTVT
6236.	TTVTD
6237.	TVTDL
6238.	VTDLE
6239.	TDLEA
6240.	DLEAA
6241.	LEAAK
6242.	EAAKQ
6243.	AAKQQ
6244.	AKQQL
6245.	KQQLE
6246.	QQLEE
6247.	QLEEK
6248.	LEEKV
6249.	EEKVT
6250.	EKVTD
6251.	KVTDL
6252.	VTDLE
6253.	TDLES
6254.	DLESE
6255.	LESEK
6256.	ESEKQ
6257.	SEKQE
6258.	EKQEL
6259.	KQELR
6260.	QELRE
6261.	ELREE
6262.	LREEL
6263.	REELD
6264.	EELDK
6265.	ELDKA
6266.	LDKAI
6267.	DKAIE
6268.	KAIEN
6269.	AIENL
6270.	IENLD
6271.	ENLDE
6272.	NLDEM
6273.	LDEMA
6274.	DEMAY
6275.	EMAYE
6276.	MAYEA
6277.	AYEAM
6278.	YEAME
6279.	EAMEF
6280.	AMEFE
6281.	MEFEK
6282.	EFEKE
6283.	FEKEK
6284.	EKEKH
6285.	KEKHG
6286.	EKHGI
6287.	KHGIK
6288.	HGIKP
6289.	GIKPG
6290.	IKPGR
6291.	KPGRR
6292.	PGRRG
6293.	GRRGS
6294.	RRGSI

5-mer peptides of CT795 (hypothetical protein,

163 amino acids), ACCESSION# NP 220315.1,

GI: 15605529 (SEQ ID NOS: 6295-6453)

6295.	MRFLL
6296.	RFLLA
6297.	FLLAL
6298.	LLALF
6299.	LALFS
6300.	ALFSL
6301.	LFSLI
6302.	FSLIL
6303.	SLILV
6304.	LILVL
6305.	ILVLP
6306.	LVLPA
6307.	VLPAT
6308.	LPATE
6309.	PATEA
6310.	ATEAF
6311.	TEAFS
6312.	EAFST
6313.	AFSTE
6314.	FSTED
6315.	STEDK
6316.	TEDKQ
6317.	EDKQC
6318.	DKQCQ
6319.	KQCQQ
6320.	QCQQE
6321.	CQQEA
6322.	QQEAE
6323.	QEAEE
6324.	EAEED
6325.	AEEDC
6326.	EEDCS
6327.	EDCSQ
6328.	DCSQV
6329.	CSQVA
6330.	SQVAD
6331.	QVADT
6332.	VADTC
6333.	ADTCV
6334.	DTCVF
6335.	TCVFY
6336.	CVFYS
6337.	VFYSY
6338.	FYSYA
6339.	YSYAE
6340.	SYAEG
6341.	YAEGL
6342.	AEGLE
6343.	EGLEH
6344.	GLEHA
6345.	LEHAR
6346.	EHARD
6347.	HARDE
6348.	ARDEG
6349.	RDEGK
6350.	DEGKL
6351.	EGKLT
6352.	GKLTL
6353.	KLTLV
6354.	LTLVV
6355.	TLVVL
6356.	LVVLL
6357.	VVLLD
6358.	VLLDT
6359.	LLDTS
6360.	LDTSG
6361.	DTSGY
6362.	TSGYS
6363.	SGYSF
6364.	GYSFE
6365.	YSFET
6366.	SFETL
6367.	FETLA
6368.	ETLAD
6369.	TLADA
6370.	LADAA
6371.	ADAAH
6372.	DAAHA
6373.	AAHAM
6374.	AHAME
6375.	HAMES
6376.	AMESS
6377.	MESSL
6378.	ESSLL
6379.	SSLLS
6380.	SLLST
6381.	LLSTF
6382.	LSTFA
6383.	STFAD
6384.	TFADF
6385.	FADFV
6386.	ADFVV
6387.	DFVVL
6388.	FVVLS
6389.	VVLSR
6390.	VLSRR
6391.	LSRRE
6392.	SRREA
6393.	RREAV
6394.	REAVP
6395.	EAVPL
6396.	AVPLI
6397.	VPLIY
6398.	PLIYP
6399.	LIYPP
6400.	IYPPV
6401.	YPPVP
6402.	PPVPD
6403.	PVPDP
6404.	VPDPM
6405.	PDPMV
6406.	DPMVG
6407.	PMVGE
6408.	MVGEI
6409.	VGEIA
6410.	GEIAL
6411.	EIALF
6412.	IALFL
6413.	ALFLE
6414.	LFLEA
6415.	FLEAF
6416.	LEAFS
6417.	EAFSD
6418.	AFSDQ
6419.	FSDQT
6420.	SDQTF
6421.	DQTFP
6422.	QTFPS
6423.	TFPSQ
6424.	FPSQP
6425.	PSQPV
6426.	SQPVI
6427.	QPVIV
6428.	PVIVT
6429.	VIVTL
6430.	IVTLA
6431.	VTLAI
6432.	TLAIG
6433.	LAIGA
6434.	AIGAS
6435.	IGASS
6436.	GASSA
6437.	ASSAE
6438.	SSAEI
6439.	SAEIM
6440.	AEIMD
6441.	EIMDI
6442.	IMDIT
6443.	MDITE
6444.	DITEI
6445.	ITEIP
6446.	TEIPS
6447.	EIPSI
6448.	IPSIN
6449.	PSINP
6450.	SINPE
6451.	INPEF
6452.	NPEFV
6453.	PEFVE

5-mer peptides of CT621 (hypothetical protein,

832 amino acids), ACCESSION# NP 220138.1,

GI: 15605352 (SEQ ID NOS: 6454-7281)

6454.	MNRIH
6455.	NRIHR
6456.	RIHRT
6457.	IHRTQ
6458.	HRTQG
6459.	RTQGS
6460.	TQGSL
6461.	QGSLT
6462.	GSLTD
6463.	SLTDY
6464.	LTDYN
6465.	TDYNS
6466.	DYNST
6467.	YNSTL
6468.	NSTLE
6469.	STLEA
6470.	TLEAI
6471.	LEAIA
6472.	EAIAK
6473.	AIAKK
6474.	IAKKI
6475.	AKKIA
6476.	KKIAK
6477.	KIAKP
6478.	IAKPD
6479.	AKPDS
6480.	KPDSA
6481.	PDSAT
6482.	DSATI
6483.	SATIV
6484.	ATIVS
6485.	TIVSQ
6486.	IVSQV
6487.	VSQVA
6488.	SQVAQ
6489.	QVAQY
6490.	VAQYE
6491.	AQYEQ
6492.	QYEQF
6493.	YEQFK
6494.	EQFKM
6495.	QFKME
6496.	FKMEQ
6497.	KMEQE
6498.	MEQEA
6499.	EQEAL
6500.	QEALK
6501.	EALKA
6502.	ALKAL
6503.	LKALL
6504.	KALLV
6505.	ALLVS
6506.	LLVSF
6507.	LVSFD
6508.	VSFDQ
6509.	SFDQK
6510.	FDQKA
6511.	DQKAD
6512.	QKADQ
6513.	KADQR
6514.	ADQRY
6515.	DQRYR
6516.	QRYRN
6517.	RYRNL
6518.	YRNLI
6519.	RNLIQ
6520.	NLIQR
6521.	LIQRL
6522.	IQRLE
6523.	QRLEQ
6524.	RLEQL
6525.	LEQLD
6526.	EQLDV
6527.	QLDVD
6528.	LDVDR
6529.	DVDRQ
6530.	VDRQT
6531.	DRQTG
6532.	RQTGR
6533.	QTGRS
6534.	TGRST
6535.	GRSTE
6536.	RSTES
6537.	STESQ
6538.	TESQH
6539.	ESQHI
6540.	SQHIQ
6541.	QHIQE
6542.	HIQEK
6543.	IQEKP
6544.	QEKPM
6545.	EKPMA
6546.	KPMAS
6547.	PMASL
6548.	MASLQ
6549.	ASLQS
6550.	SLQSE
6551.	LQSEN
6552.	QSENQ
6553.	SENQV
6554.	ENQVV
6555.	NQVVA
6556.	QVVAQ
6557.	VVAQA
6558.	VAQAV
6559.	AQAVV
6560.	QAVVQ
6561.	AVVQS
6562.	VVQSD
6563.	VQSDS
6564.	QSDSS
6565.	SDSSM
6566.	DSSMP
6567.	SSMPI
6568.	SMPIF
6569.	MPIFT
6570.	PIFTG
6571.	IFTGI
6572.	FTGIK
6573.	TGIKQ
6574.	GIKQS
6575.	IKQSW
6576.	KQSWA
6577.	QSWAV
6578.	SWAVR
6579.	WAVRL
6580.	AVRLV
6581.	VRLVQ
6582.	RLVQG
6583.	LVQGI
6584.	VQGIR
6585.	QGIRE
6586.	GIREI
6587.	IREIL
6588.	REILD
6589.	EILDQ
6590.	ILDQV
6591.	LDQVL
6592.	DQVLV
6593.	QVLVD
6594.	VLVDT
6595.	LVDTS
6596.	VDTSL
6597.	DTSLF
6598.	TSLFT
6599.	SLFTE
6600.	LFTEE
6601.	FTEEE
6602.	TEEER
6603.	EEERG
6604.	EERGD
6605.	ERGDL
6606.	RGDLL
6607.	GDLLA
6608.	DLLAI
6609.	LLAIR
6610.	LAIRM
6611.	AIRMD
6612.	IRMDA
6613.	RMDAA
6614.	MDAAS
6615.	DAASL
6616.	AASLQ
6617.	ASLQD
6618.	SLQDK
6619.	LQDKQ
6620.	QDKQE
6621.	DKQER
6622.	KQERL
6623.	QERLS
6624.	ERLST
6625.	RLSTE
6626.	LSTED
6627.	STEDI
6628.	TEDIR
6629.	EDIRS
6630.	DIRSL
6631.	IRSLL
6632.	RSLLS
6633.	SLLSL
6634.	LLSLS
6635.	LSLSN
6636.	SLSND
6637.	LSNDV
6638.	SNDVM
6639.	NDVMR
6640.	DVMRV
6641.	VMRVL
6642.	MRVLQ
6643.	RVLQK
6644.	VLQKA
6645.	LQKAS
6646.	QKASV
6647.	KASVS
6648.	ASVSS
6649.	SVSST
6650.	VSSTR
6651.	SSTRQ
6652.	STRQL
6653.	TRQLE
6654.	RQLEL
6655.	QLELI
6656.	LELIQ
6657.	ELIQS
6658.	LIQSL
6659.	IQSLI
6660.	QSLID
6661.	SLIDI
6662.	LIDIF
6663.	IDIFG
6664.	DIFGT
6665.	IFGTE
6666.	FGTEE
6667.	GTEEN
6668.	TEENL
6669.	EENLE
6670.	ENLEQ
6671.	NLEQS
6672.	LEQSF
6673.	EQSFA
6674.	QSFAQ
6675.	SFAQV
6676.	FAQVR
6677.	AQVRL
6678.	QVRLE
6679.	VRLEN
6680.	RLENF
6681.	LENFQ
6682.	ENFQA
6683.	NFQAI
6684.	FQAIL
6685.	QAILS
6686.	AILSV
6687.	ILSVI
6688.	LSVIK
6689.	SVIKE
6690.	VIKER
6691.	IKERL
6692.	KERLT
6693.	ERLTE
6694.	RLTEE
6695.	LTEEE
6696.	TEEEF
6697.	EEEFR
6698.	EEFRV
6699.	EFRVF
6700.	FRVFQ
6701.	RVFQE
6702.	VFQEV
6703.	FQEVS
6704.	QEVSE
6705.	EVSEE
6706.	VSEEI
6707.	SEEIS
6708.	EEISS
6709.	EISSI
6710.	ISSIQ
6711.	SSIQR
6712.	SIQRT
6713.	IQRTS
6714.	QRTSE
6715.	RTSES
6716.	TSESH
6717.	SESHL
6718.	ESHLS
6719.	SHLSP
6720.	HLSPE
6721.	LSPEH
6722.	SPEHI
6723.	PEHIE
6724.	EHIEA
6725.	HIEAI
6726.	IEAIA
6727.	EAIAR
6728.	AIARV
6729.	IARVG
6730.	ARVGG
6731.	RVGGH
6732.	VGGHL
6733.	GGHLS
6734.	GHLSA
6735.	HLSAK
6736.	LSAKI
6737.	SAKIV
6738.	AKIVE
6739.	KIVES
6740.	IVESE
6741.	VESEL
6742.	ESELK
6743.	SELKA
6744.	ELKAS
6745.	LKASQ
6746.	KASQK
6747.	ASQKV
6748.	SQKVD
6749.	QKVDL
6750.	KVDLC
6751.	VDLCQ
6752.	DLCQR
6753.	LCQRI
6754.	CQRIA
6755.	QRIAA
6756.	RIAAM
6757.	IAAMY
6758.	AAMYQ
6759.	AMYQE
6760.	MYQEQ
6761.	YQEQV
6762.	QEQVD
6763.	EQVDA
6764.	QVDAV
6765.	VDAVQ
6766.	DAVQA
6767.	AVQAY
6768.	VQAYH
6769.	QAYHS
6770.	AYHSL
6771.	YHSLE
6772.	HSLEQ
6773.	SLEQD
6774.	LEQDA
6775.	EQDAL
6776.	QDALF
6777.	DALFV
6778.	ALFVN
6779.	LFVNS
6780.	FVNSR
6781.	VNSRQ
6782.	NSRQH
6783.	SRQHS
6784.	RQHSH
6785.	QHSHF
6786.	HSHFV
6787.	SHFVQ
6788.	HFVQV
6789.	FVQVI
6790.	VQVIS
6791.	QVISL
6792.	VISLV
6793.	ISLVS
6794.	SLVSS
6795.	LVSSL
6796.	VSSLM
6797.	SSLMH
6798.	SLMHS
6799.	LMHSL
6800.	MHSLS
6801.	HSLSP
6802.	SLSPT
6803.	LSPTS
6804.	SPTSE
6805.	PTSEE
6806.	TSEEE
6807.	SEEER
6808.	EEERI
6809.	EERIL
6810.	ERILL
6811.	RILLN
6812.	ILLNP
6813.	LLNPA
6814.	LNPAM
6815.	NPAMM
6816.	PAMMV
6817.	AMMVS
6818.	MMVSV
6819.	MVSVL
6820.	VSVLP
6821.	SVLPT
6822.	VLPTV
6823.	LPTVR
6824.	PTVRA
6825.	TVRAI
6826.	VRAIG
6827.	RAIGL
6828.	AIGLR
6829.	IGLRF
6830.	GLRFD
6831.	LRFDF
6832.	RFDFL
6833.	FDFLT
6834.	DFLTA
6835.	FLTAE
6836.	LTAEQ
6837.	TAEQQ
6838.	AEQQQ
6839.	EQQQM
6840.	QQQMV
6841.	QQMVN
6842.	QMVNA
6843.	MVNAA
6844.	VNAAV
6845.	NAAVS
6846.	AAVSS
6847.	AVSSL
6848.	VSSLQ
6849.	SSLQQ
6850.	SLQQQ
6851.	LQQQQ
6852.	QQQQL
6853.	QQQLD
6854.	QQLDE
6855.	QLDEF
6856.	LDEFL
6857.	DEFLG
6858.	EFLGV
6859.	FLGVL
6860.	LGVLC
6861.	GVLCA
6862.	VLCAH
6863.	LCAHL
6864.	CAHLV
6865.	AHLVV
6866.	HLVVV
6867.	LVVVN
6868.	VVVNC
6869.	VVNCQ
6870.	VNCQN
6871.	NCQNK
6872.	CQNKE
6873.	QNKET
6874.	NKETG
6875.	KETGL
6876.	ETGLL
6877.	TGLLE
6878.	GLLEG
6879.	LLEGL
6880.	LEGLE
6881.	EGLEE
6882.	GLEES
6883.	LEESF
6884.	EESFS
6885.	ESFSE
6886.	SFSET
6887.	FSETL
6888.	SETLS
6889.	ETLSG
6890.	TLSGL
6891.	LSGLS
6892.	SGLSN
6893.	GLSNN
6894.	LSNNF
6895.	SNNFV
6896.	NNFVL
6897.	NFVLT
6898.	FVLTA
6899.	VLTAK
6900.	LTAKM
6901.	TAKMQ
6902.	AKMQD
6903.	KMQDI
6904.	MQDIL
6905.	QDILQ
6906.	DILQV
6907.	ILQVC
6908.	LQVCS
6909.	QVCSL
6910.	VCSLQ
6911.	CSLQG
6912.	SLQGF
6913.	LQGFV
6914.	QGFVT
6915.	GFVTL
6916.	FVTLA
6917.	VTLAN
6918.	TLANG
6919.	LANGD
6920.	ANGDR
6921.	NGDRY
6922.	GDRYE
6923.	DRYEL
6924.	RYELF
6925.	YELFS
6926.	ELFSY
6927.	LFSYN
6928.	FSYND
6929.	SYNDS
6930.	YNDSG
6931.	NDSGE
6932.	DSGEA
6933.	SGEAV
6934.	GEAVC
6935.	EAVCD
6936.	AVCDE
6937.	VCDEI
6938.	CDEIA
6939.	DEIAL
6940.	EIALG
6941.	IALGD
6942.	ALGDG
6943.	LGDGF
6944.	GDGFH
6945.	DGFHK
6946.	GFHKV
6947.	FHKVL
6948.	HKVLG
6949.	KVLGT
6950.	VLGTM
6951.	LGTML
6952.	GTMLA
6953.	TMLAV
6954.	MLAVA
6955.	LAVAL
6956.	AVALS
6957.	VALSQ
6958.	ALSQA
6959.	LSQAE
6960.	SQAEV
6961.	QAEVF
6962.	AEVFK
6963.	EVFKQ
6964.	VFKQE
6965.	FKQEC
6966.	KQECD
6967.	QECDR
6968.	ECDRF
6969.	CDRFI
6970.	DRFIL
6971.	RFILQ
6972.	FILQA
6973.	ILQAD
6974.	LQADS
6975.	QADSE
6976.	ADSEK
6977.	DSEKN
6978.	SEKNM
6979.	EKNMI
6980.	KNMIH
6981.	NMIHK
6982.	MIHKR
6983.	IHKRM
6984.	HKRMV
6985.	KRMVQ
6986.	RMVQG
6987.	MVQGE
6988.	VQGEQ
6989.	QGEQK
6990.	GEQKS
6991.	EQKSL
6992.	QKSLF
6993.	KSLFL
6994.	SLFLT
6995.	LFLTK
6996.	FLTKM
6997.	LTKMQ
6998.	TKMQT
6999.	KMQTE
7000.	MQTEL
7001.	QTELN
7002.	TELNA
7003.	ELNAG
7004.	LNAGK
7005.	NAGKT
7006.	AGKTI
7007.	GKTIA
7008.	KTIAQ
7009.	TIAQT
7010.	IAQTK
7011.	AQTKE
7012.	QTKEV
7013.	TKEVE
7014.	KEVEA
7015.	EVEAS
7016.	VEASP
7017.	EASPL
7018.	ASPLP
7019.	SPLPS
7020.	PLPSA
7021.	LPSAV
7022.	PSAVA
7023.	SAVAS
7024.	AVASV
7025.	VASVL
7026.	ASVLI
7027.	SVLID
7028.	VLIDH
7029.	LIDHY
7030.	IDHYM
7031.	DHYMP
7032.	HYMPK
7033.	YMPKE
7034.	MPKEV
7035.	PKEVE
7036.	KEVEF
7037.	EVEFL
7038.	VEFLE
7039.	EFLEK
7040.	FLEKI
7041.	LEKIS
7042.	EKISS
7043.	KISSR
7044.	ISSRL
7045.	SSRLY
7046.	SRLYY
7047.	RLYYG
7048.	LYYGN
7049.	YYGNK
7050.	YGNKG
7051.	GNKGS
7052.	NKGSD
7053.	KGSDI
7054.	GSDIG
7055.	SDIGN
7056.	DIGNT
7057.	IGNTI
7058.	GNTIL
7059.	NTILD
7060.	TILDA
7061.	ILDAI
7062.	LDAIS
7063.	DAISL
7064.	AISLY
7065.	ISLYV
7066.	SLYVN
7067.	LYVNS
7068.	YVNSA
7069.	VNSAT
7070.	NSATY
7071.	SATYF
7072.	ATYFG
7073.	TYFGF
7074.	YFGFA
7075.	FGFAN
7076.	GFANY
7077.	FANYI
7078.	ANYIG
7079.	NYIGQ
7080.	YIGQP
7081.	IGQPP
7082.	GQPPV
7083.	QPPVV
7084.	PPVVG
7085.	PVVGK
7086.	VVGKT
7087.	VGKTR
7088.	GKTRE
7089.	KTREN
7090.	TRENT
7091.	RENIF
7092.	ENIFA
7093.	NIFAG
7094.	IFAGS
7095.	FAGSA
7096.	AGSAD
7097.	GSADN
7098.	SADNA
7099.	ADNAK
7100.	DNAKA
7101.	NAKAK
7102.	AKAKL
7103.	KAKLD
7104.	AKLDE
7105.	KLDEE
7106.	LDEEK
7107.	DEEKK
7108.	EEKKQ
7109.	EKKQV
7110.	KKQVD
7111.	KQVDV
7112.	QVDVF
7113.	VDVFL
7114.	DVFLE
7115.	VFLEI
7116.	FLEIT
7117.	LEITE
7118.	EITEA
7119.	ITEAA
7120.	TEAAK
7121.	EAAKT
7122.	AAKTT
7123.	AKTTV
7124.	KTTVT
7125.	TTVTN
7126.	TVTNQ
7127.	VTNQQ
7128.	TNQQS
7129.	NQQSA
7130.	QQSAV
7131.	QSAVT
7132.	SAVTN
7133.	AVTND
7134.	VTNDD
7135.	TNDDK
7136.	NDDKL
7137.	DDKLS
7138.	DKLST
7139.	KLSTE
7140.	LSTEQ
7141.	STEQK
7142.	TEQKA
7143.	EQKAK
7144.	QKAKI
7145.	KAKIK
7146.	AKIKA
7147.	KIKAE
7148.	IKAEL
7149.	KAELT
7150.	AELTQ
7151.	ELTQY
7152.	LTQYT
7153.	TQYTD
7154.	QYTDM
7155.	YTDML
7156.	TDMLN
7157.	DMLNA
7158.	MLNAI
7159.	LNAIS
7160.	NAISN
7161.	AISNS
7162.	ISNSL
7163.	SNSLT
7164.	NSLTS
7165.	SLTSL
7166.	LTSLK
7167.	TSLKT
7168.	SLKTQ
7169.	LKTQL
7170.	KTQLA
7171.	TQLAP
7172.	QLAPL
7173.	LAPLS
7174.	APLSV
7175.	PLSVS
7176.	LSVST
7177.	SVSTV
7178.	VSTVE
7179.	STVEG
7180.	TVEGV
7181.	VEGVD
7182.	EGVDG
7183.	GVDGV
7184.	VDGVF
7185.	DGVFE
7186.	GVFEV
7187.	VFEVK
7188.	FEVKN
7189.	EVKNG
7190.	VKNGI
7191.	KNGIP
7192.	NGIPG
7193.	GIPGE
7194.	IPGEN
7195.	PGENG
7196.	GENGK
7197.	ENGKN
7198.	NGKNW
7199.	GKNWR
7200.	KNWRL
7201.	NWRLV
7202.	WRLVL
7203.	RLVLQ
7204.	LVLQT
7205.	VLQTL
7206.	LQTLE
7207.	QTLED
7208.	TLEDT
7209.	LEDTV
7210.	EDTVV
7211.	DTVVS
7212.	TVVSG
7213.	VVSGE
7214.	VSGEV
7215.	SGEVG
7216.	GEVGS
7217.	EVGSP
7218.	VGSPT
7219.	GSPTN
7220.	SPTNI
7221.	PTNIG
7222.	TNIGM
7223.	NIGMF
7224.	IGMFQ
7225.	GMFQM
7226.	MFQMQ
7227.	FQMQA
7228.	QMQAL
7229.	MQALV
7230.	QALVH
7231.	ALVHL
7232.	LVHLN
7233.	VHLNQ
7234.	HLNQQ
7235.	LNQQA
7236.	NQQAY
7237.	QQAYA
7238.	QAYAD
7239.	AYADM
7240.	YADMG
7241.	ADMGQ
7242.	DMGQN
7243.	MGQNF
7244.	GQNFQ
7245.	QNFQL
7246.	NFQLE
7247.	FQLEL
7248.	QLELQ
7249.	LELQM
7250.	ELQMH
7251.	LQMHL
7252.	QMHLT
7253.	MHLTS
7254.	HLTSM
7255.	LTSMQ
7256.	TSMQQ
7257.	SMQQE
7258.	MQQEW
7259.	QQEWM
7260.	QEWMV
7261.	EWMVV
7262.	WMVVA
7263.	MVVAT
7264.	VVATS
7265.	VATSL
7266.	ATSLQ
7267.	TSLQL
7268.	SLQLL
7269.	LQLLN
7270.	QLLNQ
7271.	LLNQI
7272.	LNQIY
7273.	NQIYL
7274.	QIYLG
7275.	IYLGL
7276.	YLGLA
7277.	LGLAR
7278.	GLARN
7279.	LARNL
7280.	ARNLL
7281.	RNLLR

5-mer peptides of CT622 (hypothetical protein,

647 amino acids), ACCESSION# NP 220139.1,

GI: 15605353 (SEQ ID NOS: 7282-7924)

7282.	MESGP
7283.	ESGPE
7284.	SGPES
7285.	GPESV
7286.	PESVS
7287.	ESVSS
7288.	SVSSN
7289.	VSSNQ
7290.	SSNQS
7291.	SNQSS
7292.	NQSSM
7293.	QSSMN
7294.	SSMNP
7295.	SMNPI
7296.	MNPII
7297.	NPIIN
7298.	PIING
7299.	IINGQ
7300.	INGQI
7301.	NGQIA
7302.	GQIAS
7303.	QIASN
7304.	IASNS
7305.	ASNSE
7306.	SNSET
7307.	NSETK
7308.	SETKE
7309.	ETKES
7310.	TKEST
7311.	KESTK
7312.	ESTKE
7313.	STKES
7314.	TKESE
7315.	KESEA
7316.	ESEAS
7317.	SEASP
7318.	EASPS
7319.	ASPSA
7320.	SPSAS
7321.	PSASS
7322.	SASSS
7323.	ASSSV
7324.	SSSVS
7325.	SSVSS
7326.	SVSSW
7327.	VSSWS
7328.	SSWSF
7329.	SWSFL
7330.	WSFLS
7331.	SFLSS
7332.	FLSSA
7333.	LSSAK
7334.	SSAKH
7335.	SAKHA
7336.	AKHAL
7337.	KHALI
7338.	HALIS
7339.	ALISL
7340.	LISLR
7341.	ISLRD
7342.	SLRDA
7343.	LRDAI
7344.	RDAIL
7345.	DAILN
7346.	AILNK
7347.	ILNKN
7348.	LNKNS
7349.	NKNSS
7350.	KNSSP
7351.	NSSPT
7352.	SSPTD
7353.	SPTDS
7354.	PTDSL
7355.	TDSLS
7356.	DSLSQ
7357.	SLSQL
7358.	LSQLE
7359.	SQLEA
7360.	QLEAS
7361.	LEAST
7362.	EASTS
7363.	ASTST
7364.	STSTS
7365.	TSTST
7366.	STSTV
7367.	TSTVT
7368.	STVTR
7369.	TVTRV
7370.	VTRVA
7371.	TRVAA
7372.	RVAAR
7373.	VAARD
7374.	AARDY
7375.	ARDYN
7376.	RDYNE
7377.	DYNEA
7378.	YNEAK
7379.	NEAKS
7380.	EAKSN
7381.	AKSNF
7382.	KSNFD
7383.	SNFDT
7384.	NFDTA
7385.	FDTAK
7386.	DTAKS
7387.	TAKSG
7388.	AKSGL
7389.	KSGLE
7390.	SGLEN
7391.	GLENA
7392.	LENAT
7393.	ENATT
7394.	NATTL
7395.	ATTLA
7396.	TTLAE
7397.	TLAEY
7398.	LAEYE
7399.	AEYET
7400.	EYETK
7401.	YETKM
7402.	ETKMA
7403.	TKMAD
7404.	KMADL
7405.	MADLM
7406.	ADLMA
7407.	DLMAA
7408.	LMAAL
7409.	MAALQ
7410.	AALQD
7411.	ALQDM
7412.	LQDME
7413.	QDMER
7414.	DMERL
7415.	MERLA
7416.	ERLAK
7417.	RLAKQ
7418.	LAKQK
7419.	AKQKA
7420.	KQKAE
7421.	QKAEV
7422.	KAEVT
7423.	AEVTR
7424.	EVTRI
7425.	VTRIK
7426.	TRIKE
7427.	RIKEA
7428.	IKEAL
7429.	KEALQ
7430.	EALQE
7431.	ALQEK
7432.	LQEKQ
7433.	QEKQE
7434.	EKQEV
7435.	KQEVI
7436.	QEVID
7437.	EVIDK
7438.	VIDKL
7439.	IDKLN
7440.	DKLNQ
7441.	KLNQL
7442.	LNQLV
7443.	NQLVK
7444.	QLVKL
7445.	LVKLE
7446.	VKLEK
7447.	KLEKQ
7448.	LEKQN
7449.	EKQNQ
7450.	KQNQT
7451.	QNQTL
7452.	NQTLK
7453.	QTLKE
7454.	TLKET
7455.	LKETL
7456.	KETLT
7457.	ETLTT
7458.	TLTTT
7459.	LTTTD
7460.	TTTDS
7461.	TTDSA
7462.	TDSAD
7463.	DSADQ
7464.	SADQI
7465.	ADQIP
7466.	DQIPA
7467.	QIPAI
7468.	IPAIN
7469.	PAINS
7470.	AINSQ
7471.	INSQL
7472.	NSQLE
7473.	SQLEI
7474.	QLEIN
7475.	LEINK
7476.	EINKN
7477.	INKNS
7478.	NKNSA
7479.	KNSAD
7480.	NSADQ
7481.	SADQI
7482.	ADQII
7483.	DQIIK
7484.	QIIKD
7485.	IIKDL
7486.	IKDLE
7487.	KDLEG
7488.	DLEGQ
7489.	LEGQN
7490.	EGQNI
7491.	GQNIS
7492.	QNISY
7493.	NISYE
7494.	ISYEA
7495.	SYEAV
7496.	YEAVL
7497.	EAVLT
7498.	AVLTN
7499.	VLTNA
7500.	LTNAG
7501.	TNAGE
7502.	NAGEV
7503.	AGEVI
7504.	GEVIK
7505.	EVIKA
7506.	VIKAS
7507.	IKASS
7508.	KASSE
7509.	ASSEA
7510.	SSEAG
7511.	SEAGI
7512.	EAGIK
7513.	AGIKL
7514.	GIKLG
7515.	IKLGQ
7516.	KLGQA
7517.	LGQAL
7518.	GQALQ
7519.	QALQS
7520.	ALQSI
7521.	LQSIV
7522.	QSIVD
7523.	SIVDA
7524.	IVDAG
7525.	VDAGD
7526.	DAGDQ
7527.	AGDQS
7528.	GDQSQ
7529.	DQSQA
7530.	QSQAA
7531.	SQAAV
7532.	QAAVL
7533.	AAVLQ
7534.	AVLQA
7535.	VLQAQ
7536.	LQAQQ
7537.	QAQQN
7538.	AQQNN
7539.	QQNNS
7540.	QNNSP
7541.	NNSPD
7542.	NSPDN
7543.	SPDNI
7544.	PDNIA
7545.	DNIAA
7546.	NIAAT
7547.	IAATK
7548.	AATKK
7549.	ATKKL
7550.	TKKLI
7551.	KKLID
7552.	KLIDA
7553.	LIDAA
7554.	IDAAE
7555.	DAAET
7556.	AAETK
7557.	AETKV
7558.	ETKVN
7559.	TKVNE
7560.	KVNEL
7561.	VNELK
7562.	NELKQ
7563.	ELKQE
7564.	LKQEH
7565.	KQEHT
7566.	QEHTG
7567.	EHTGL
7568.	HTGLT
7569.	TGLTD
7570.	GLTDS
7571.	LTDSP
7572.	TDSPL
7573.	DSPLV
7574.	SPLVK
7575.	PLVKK
7576.	LVKKA
7577.	VKKAE
7578.	KKAEE
7579.	KAEEQ
7580.	AEEQI
7581.	EEQIS
7582.	EQISQ
7583.	QISQA
7584.	ISQAQ
7585.	SQAQK
7586.	QAQKD
7587.	AQKDI
7588.	QKDIQ
7589.	KDIQE
7590.	DIQEI
7591.	IQEIK
7592.	QEIKP
7593.	EIKPS
7594.	IKPSG
7595.	KPSGS
7596.	PSGSD
7597.	SGSDI
7598.	GSDIP
7599.	SDIPI
7600.	DIPIV
7601.	IPIVG
7602.	PIVGP
7603.	IVGPS
7604.	VGPSG
7605.	GPSGS
7606.	PSGSA
7607.	SGSAA
7608.	GSAAS
7609.	SAASA
7610.	AASAG
7611.	ASAGS
7612.	SAGSA
7613.	AGSAV
7614.	GSAVG
7615.	SAVGA
7616.	AVGAL
7617.	VGALK
7618.	GALKS
7619.	ALKSS
7620.	LKSSN
7621.	KSSNN
7622.	SSNNS
7623.	SNNSG
7624.	NNSGR
7625.	NSGRI
7626.	SGRIS
7627.	GRISL
7628.	RISLL
7629.	ISLLL
7630.	SLLLD
7631.	LLLDD
7632.	LLDDV
7633.	LDDVD
7634.	DDVDN
7635.	DVDNE
7636.	VDNEM
7637.	DNEMA
7638.	NEMAA
7639.	EMAAI
7640.	MAAIA
7641.	AAIAM
7642.	AIAMQ
7643.	IAMQG
7644.	AMQGF
7645.	MQGFR
7646.	QGFRS
7647.	GFRSM
7648.	FRSMI
7649.	RSMIE
7650.	SMIEQ
7651.	MIEQF
7652.	IEQFN
7653.	EQFNV
7654.	QFNVN
7655.	FNVNN
7656.	NVNNP
7657.	VNNPA
7658.	NNPAT
7659.	NPATA
7660.	PATAK
7661.	ATAKE
7662.	TAKEL
7663.	AKELQ
7664.	KELQA
7665.	ELQAM
7666.	LQAME
7667.	QAMEA
7668.	AMEAQ
7669.	MEAQL
7670.	EAQLT
7671.	AQLTA
7672.	QLTAM
7673.	LTAMS
7674.	TAMSD
7675.	AMSDQ
7676.	MSDQL
7677.	SDQLV
7678.	DQLVG
7679.	QLVGA
7680.	LVGAD
7681.	VGADG
7682.	GADGE
7683.	ADGEL
7684.	DGELP
7685.	GELPA
7686.	ELPAE
7687.	LPAEI
7688.	PAEIQ
7689.	AEIQA
7690.	EIQAI
7691.	IQAIK
7692.	QAIKD
7693.	AIKDA
7694.	IKDAL
7695.	KDALA
7696.	DALAQ
7697.	ALAQA
7698.	LAQAL
7699.	AQALK
7700.	QALKQ
7701.	ALKQP
7702.	LKQPS
7703.	KQPST
7704.	QPSTD
7705.	PSTDG
7706.	STDGL
7707.	TDGLA
7708.	DGLAT
7709.	GLATA
7710.	LATAM
7711.	ATAMG
7712.	TAMGQ
7713.	AMGQV
7714.	MGQVA
7715.	GQVAF
7716.	QVAFA
7717.	VAFAA
7718.	AFAAA
7719.	FAAAK
7720.	AAAKV
7721.	AAKVG
7722.	AKVGG
7723.	KVGGG
7724.	VGGGS
7725.	GGGSA
7726.	GGSAG
7727.	GSAGT
7728.	SAGTA
7729.	AGTAG
7730.	GTAGT
7731.	TAGTV
7732.	AGTVQ
7733.	GTVQM
7734.	TVQMN
7735.	VQMNV
7736.	QMNVK
7737.	MNVKQ
7738.	NVKQL
7739.	VKQLY
7740.	KQLYK
7741.	QLYKT
7742.	LYKTA
7743.	YKTAF
7744.	KTAFS
7745.	TAFSS
7746.	AFSST
7747.	FSSTS
7748.	SSTSS
7749.	STSSS
7750.	TSSSS
7751.	SSSSY
7752.	SSSYA
7753.	SSYAA
7754.	SYAAA
7755.	YAAAL
7756.	AAALS
7757.	AALSD
7758.	ALSDG
7759.	LSDGY
7760.	SDGYS
7761.	DGYSA
7762.	GYSAY
7763.	YSAYK
7764.	SAYKT
7765.	AYKTL
7766.	YKTLN
7767.	KTLNS
7768.	TLNSL
7769.	LNSLY
7770.	NSLYS
7771.	SLYSE
7772.	LYSES
7773.	YSESR
7774.	SESRS
7775.	ESRSG
7776.	SRSGV
7777.	RSGVQ
7778.	SGVQS
7779.	GVQSA
7780.	VQSAI
7781.	QSAIS
7782.	SAISQ
7783.	AISQT
7784.	ISQTA
7785.	SQTAN
7786.	QTANP
7787.	TANPA
7788.	ANPAL
7789.	NPALS
7790.	PALSR
7791.	ALSRS
7792.	LSRSV
7793.	SRSVS
7794.	RSVSR
7795.	SVSRS
7796.	VSRSG
7797.	SRSGI
7798.	RSGIE
7799.	SGIES
7800.	GIESQ
7801.	IESQG
7802.	ESQGR
7803.	SQGRS
7804.	QGRSA
7805.	GRSAD
7806.	RSADA
7807.	SADAS
7808.	ADASQ
7809.	DASQR
7810.	ASQRA
7811.	SQRAA
7812.	QRAAE
7813.	RAAET
7814.	AAETI
7815.	AETIV
7816.	ETIVR
7817.	TIVRD
7818.	IVRDS
7819.	VRDSQ
7820.	RDSQT
7821.	DSQTL
7822.	SQTLG
7823.	QTLGD
7824.	TLGDV
7825.	LGDVY
7826.	GDVYS
7827.	DVYSR
7828.	VYSRL
7829.	YSRLQ
7830.	SRLQV
7831.	RLQVL
7832.	LQVLD
7833.	QVLDS
7834.	VLDSL
7835.	LDSLM
7836.	DSLMS
7837.	SLMST
7838.	LMSTI
7839.	MSTIV
7840.	STIVS
7841.	TIVSN
7842.	IVSNP
7843.	VSNPQ
7844.	SNPQV
7845.	NPQVN
7846.	PQVNQ
7847.	QVNQE
7848.	VNQEE
7849.	NQEEI
7850.	QEEIM
7851.	EEIMQ
7852.	EIMQK
7853.	IMQKL
7854.	MQKLT
7855.	QKLTA
7856.	KLTAS
7857.	LTASI
7858.	TASIS
7859.	ASISK
7860.	SISKA
7861.	ISKAP
7862.	SKAPQ
7863.	KAPQF
7864.	APQFG
7865.	PQFGY
7866.	QFGYP
7867.	FGYPA
7868.	GYPAV
7869.	YPAVQ
7870.	PAVQN
7871.	AVQNS
7872.	VQNSA
7873.	QNSAD
7874.	NSADS
7875.	SADSL
7876.	ADSLQ
7877.	DSLQK
7878.	SLQKF
7879.	LQKFA
7880.	QKFAA
7881.	KFAAQ
7882.	FAAQL
7883.	AAQLE
7884.	AQLER
7885.	QLERE
7886.	LEREF
7887.	EREFV
7888.	REFVD
7889.	EFVDG
7890.	FVDGE
7891.	VDGER
7892.	DGERS
7893.	GERSL
7894.	ERSLA
7895.	RSLAE
7896.	SLAES
7897.	LAESR
7898.	AESRE
7899.	ESREN
7900.	SRENA
7901.	RENAF
7902.	ENAFR
7903.	NAFRK
7904.	AFRKQ
7905.	FRKQP
7906.	RKQPA
7907.	KQPAF
7908.	QPAFI
7909.	PAFIQ
7910.	AFIQQ
7911.	FIQQV
7912.	IQQVL
7913.	QQVLV
7914.	QVLVN
7915.	VLVNI
7916.	LVNIA
7917.	VNIAS
7918.	NIASL
7919.	IASLF
7920.	ASLFS
7921.	SLFSG
7922.	LFSGY
7923.	FSGYL
7924.	SGYLS

Amino Acid Sequences of Chlamydia trachomatis Proteins
CT443, CT381, CT875, CT147, HSP60, CT376 and CT557, CT858 (CPAF), Pgp3, CT823 (cHtrA), CT681 (14014P), CT119 (IncA) and CT813 (All sequences are available from this website: www.ncbi.nlm.nih.gov/protein/15605169)

CT443 (OmcB or CRP60, 553 amino acids), ACCESSION#: NP 219955.1,
GI: 15605169
(SEQ ID NO: 7925)

1	mrigdpmnkl irravtifav tsvaslfasg vletsmaesl stnvisladt kakdntshks

61	kkarknhske tpvdrkevap vheskatgpk qdscfgrmyt vkvnddrnve itqavpeyat

121	vgspypieit atgkrdcvdv iitqqlpcea efvrsdpatt ptadgklvwk idrlgqgeks

181	kitvwvkplk egccftaatv cacpeirsvt kcgqpaicvk qegpenaclr cpvvykiniv

241	nqgtatarnv vvenpvpdgy ahssgqrvlt ftlgdmqpge hrtitvefcp lkrgratnia

301	tvsycgghkn tasvttvine pcvqvsiaga dwsyvckpve yvisvsnpgd lvlrdvvved

361	tlspgvtvle aagaqiscnk vvwtvkelnp geslqykylv raqtpgqftn nvvvkscsdc

421	gtctscaeat tywkgvaath mcvvdtcdpv cvgentvyri cvtnrgsaed tnvslmlkfs

481	kelqpvsfsg ptkgtitgnt vvfdslprlg sketvefsvt lkavsagdar geailssdtl

541	tvpvsdtent hiy

CT381 (ArtJ, 257 amino acids), ACCESSION# NP 219890.1, GI: 15605105
(SEQ ID NO: 7926)

1	mcikrkktwi aflavvcsfc ltgclkeggd snsekfivgt natyppfefv dkrgevvgfd

61	idlareisnk lgktldvref sfdalilnlk qhridavitg msitpsrlke ilmipyygee

121	ikhlvlvfkg enkhplpltq yrsvavqtgt ygeaylqsls evhirsfdst levlmevmhg

181	kspvavleps iaqvvlkdfp alstatidlp edqwvlgygi gvasdrpala lkieaavqei

241	rkegvlaele qkwglnn

CT875 (hypothetical protein, 591 amino acids), ACCESSION#: NP 219502.1,
GI: 15604718
(SEQ ID NO: 7927)

1	msirgvggng nsripshngd gsnrrsqntk gnnkvedrvc slyssrsnen respyavvdv

61	ssmiestpts gettrasrgv fsrfqrglvr vadkvrravq cawssystrr ssatraaesg

121	sssrtargas sgyreyspsa arglrlmftd fwrtrylrqt spmagvfgnl dvnearlmaa

181	ytsecadhle anklagpdgv aaareiakrw eqrvrdlqdk gaarkllndp lgrrtpnyqs

241	knpgeytvgn smfydgpqva nlqnvdtgfw ldmsnlsdvv lsreiqtglr aratleesmp

301	mlenleerfr rlqetcdaar teieesgwtr esasrmegde aqgpsraqqa fqsfvnecns

361	iefsfgsfge hvrvlcarvs rglaaageai rrcfscckgs thryaprddl spegaslaet

421	larfaddmgi ergadgtydi plvddwrrgv psiegegsds iyeimmpiye vmdmdletrr

481	sfavqqghyq dprasdydlp rasdydlprs pyptpplppr yqlqnmdvea gfreavyasf

541	vagmynyvvt qpqeripnsq qvegilrdml tngsqtfrdl mrrwnrevdr e

CT147 (hypothetical protein, 1449 amino acids), ACCESSION# NP 219650.1,
GI: 15604866
(SEQ ID NO: 7928)

1	manpstpsfn hsdlslqgrl rassqqctqa gqgdpqplsp esrgltsnfs trrdlidvve

61	esietakgse lkklriyeia lkiltiigaa ilfavplcml lgvplwipiv tcigvgiafs

121	iakgclqkrc qqireeyral hlyhryllsn kdsidgtlls rfdirfrkae eklhgldldk

181	reanhplead krydfaglah qryqvdaalg isssqdafwr gvaqqvksvk ddvvlgdkas

241	tdlypiaqqa lqaagvgfsg aagkeslldl akslsslfaw gsqvgkdshe alqqyqmrfl

301	sspilatwcg agfsasaqdf vlkgenildi asenhtkmqn aikrvqlvsv lgkmrnwkek

361	idtliqnknl dqdslrklyq diekamhkvc iedgvstsiq tqvrkvtqky lrqdlqelln

421	kkaplnesdl skmqkgissc anlvvtlles qlgtsgqtpi keveesiyrd liatilqmgs

481	aaggvtplvd gvhkairegk alrselsram slhprqsflg vqsaveklqa firdpkwgas

541	avhtsaeetl aqkqkfvsdl triqtsladw reryglfeet klnhivstdf vsrteafldt

601	lknvaeacsl eqavaelkdc edamkadlth veqkmnptei esareefkrl meelagiqeq

661	leqiaqpiye egvsgerlll ntvffhpevl rkkvqakeas lealtkgeqp sptkkktlkq

721	lsegceyfss lvskinalkt ilegsrgkki asqdirqlig ltdelalels sfqqdslesl

781	lygleglsip aasieqkkgs pksssiaekv vyashqrvhn gvkakvnrtl eafsqlikgl

841	rgslrnamit kavvaavlsv afsclaialf svqltwlpim lcvlalvlea ipsalsiwve

901	krnwkyevas lakqlvsdgr klpypdlgdq nikhlekird vygldgvael rvaeaallgv

961	qklpeeqkqe slksavkalr adakvlnkkf kklpesyqpq hsevtgvqgv teqesrddvl

1021	vaqdmaaiee lqdqyhaacl qfesystrfl aeqrkakfle ellvqkrrdv shlshqeahy

1081	tqvvshlkel ismrkgastq haskeeistk mrellslddq llkahtaqdv nrdnsingql

1141	qqqfkklsee gslqkvkall elnmclgnag qtlyhsrlkr evfeaslsgt srqllqyged

1201	lfasydgsdr sallrfvlgs gyemiseass elkslrkrwk rsasqaaiap edyekvcrvl

1261	erflkardsl rpklglplgk ssdatvglqh qirdnqrvka rvtacyqesc rnvlqhledw

1321	vrktrqesae cqkvetkire fcqkagsken laestemlfs sleedlnkip ldvlrailrs

1381	lsskvlhird qkleleklee qfaktnaivk akeaefekng evwhnqyqml ksqmeklesq

1441	krrltdkke

HSP60 (CT110, 544 amino acids), ACCESSION# AAS19616.1, GI: 42541742
(SEQ ID NO: 7929)

1	mvaknikyne earkkiqkgv ktlaeavkvt lgpkgrhvvi dksfgspqvt kdgvtvakev

61	eladkhenmg aqmvkevask tadkagdgtt tatvlaeaiy teglrnvtag anpmdlkrgi

121	dkavkvvvdq irkiskpvqh hkeiaqvati sanndaeign liaeamekvg kngsitveea

181	kgfetvldvv egmnfnrgyl ssyfatnpet qecvledalv liydkkisgi kdflpvlqqv

241	aesgrpllii aediegeala tivvnrirgg frvcavkapg fgdrrkamle diailtggql

301	iseelgmkle nanlamlgka kkvivskedt tivegmgeke alearcesik kqiedsssdy

361	dkeklqerla klsggvavir vgaateiemk ekkdrvddaq hatiaaveeg ilpgggtali

421	rciptleafl pmltnedeqi garivlkals aplkqiaana gkegaiifqq vmsrsanegy

481	dalrdaytdm leagildpak vtrsalesaa svagllltte aliaeipeek paaapampga

541	gmdy

CT376 (malate dehydrogenase, 326 amino acids) ACCESSION# NP 219885.1,
GI: 15605100
(SEQ ID NO: 7930)

1	mvsqtvsvav tggtgqiays flfslahgdv fgldcgidlr iydipgtera lsgvrmeldd

61	gafpllqrvq vttslhdafd gidaafligs vprgpgmerr dllkkngeif atqgkalntt

121	akrdakifvv gnpvntncwi amnhaprllr knfhamlrld qnrmhsmlsh raevplsavs

181	qvvvwgnhsa kqvpdftqal indrpiaeti adrdwlenim vpsvqsrgsa vieargkssa

241	asaaralaea arsiyqpkeg ewfssgvcsd hnpyglpedl ifgfpcrmla tgeyeviprl

301	pwdafirgkm qisldeilqe kasvsl

CT557 (dihydrolipoamide dehydrogenase, 465 amino acids), ACCESSION#
NP 220072.1, GI: 15605286
(SEQ ID NO: 7931)

1	mneafdcvvi gagpggyvaa itaaqaglkt aliekreagg tclnrgcips kallagaevv

61	tqirhadqfg ihvegfsiny pamvqrkdsv vrsirdglng lirsnkitvf sgrgslisst

121	evkilgenps vikahsiila tgseprafpg ipfsaespri lcstgvlnlk eipqkmaiig

181	ggvigcefas lfhtlgsevs vieassqila lnnpdisktm fdkftrqglr fvleasysni

241	edigdrvrlt ingnveeydy vlvsigrrin tenigldkag vicdergvip tdatmrtnvp

301	niyaigditg kwqlahvash qgiiaarnia ghkeeidysa vpsviftfpe vasvglspta

361	aqqqkipvkv tkfpfraigk avamgeadgf aaiishettq qilgayvigp hassliseit

421	lavrneltlp ciyetihahp tlaevwaesa llavdtplhm ppakk

CT858 (CPAF, total of 609 amino acids), ACCESSION# AAC68456.1, GI: 3329332
(SEQ ID NO: 7932)

1	mgfwrtsimk mnriwllllt fssaihspvq geslvcknal qdlsflehll qvkyapktwk

61	eqylgwdlvq ssysaqqklr tqenpstsfc qqvladfigg lndfhagvtf faiesaylpy

121	tvqkssdgrf yfvdimtfss eirvgdelle vdgapvqdvl atlygsnhkg taaeesaalr

181	tlfsrmaslg hkvpsgrttl kirrpfgttr evrvkwryvp egvgdlatia psirapqlqk

241	smrsffpkkd dafhrssslf yspmvphfwa elrnhyatsg lksgynigst dgflpvigpv

301	iweseglfra yissvtdgdg kshkvgflri ptyswqdmed fdpsgpppwe efakiiqvfs

361	sntealiidq tnnpggsvly lyallsmltd rplelpkhrm iltqdevvda ldwltllenv

421	dtnvesrlal gdnmegytvd lqvaeylksf grqvincwsk gdielstpip lfgfekihph

481	prvqyskpic vlineqdfsc adffpvvlkd ndralivgtr tagaggfvfn vqfpnrtgik

541	tcsltgslav rehgafieni gvephidlpf tandirykgy seyldkvkkl vcqlinndgt

601	iilaedgsf

Pgp3 (plasmid-encoded secretion protein, 264 amino acids), ACCESSION#
ADI51551.1, GI: 297749006
(SEQ ID NO: 7933)

1	mgnsgfylyn tencvfadni kvgqmteplk dqqiilgtks tpvaakmtas dgisltvsnn

61	sstnasitig ldaekayqli leklgnqild giadtivdst vqdildkitt dpslgllkaf

121	nnfpitnkiq cnglftpsni etllggteig kftvtpkssg smflvsadii asrmeggvvl

181	alvregdskp caisygyssg vpnlcslrts itntgltptt yslrvggles gvvwvnalsn

241	gndilgitnt snvsflevip qtna

CT823 (cHtrA, 497 amino acids), ACCESSION# NP 220344.1, GI: 15605558
(SEQ ID NO: 7934

1	mmkrllcvll stsvfsspml gysaskkdsk adiclavssg dgevsgedll kevsrgfsrv

61	aakatpgvvy ienfpktgnq aiaspgnkrg fqenpfdyfn deffnrffgl pshreqqrpq

121	qrdavrgtgf ivsedgyvvt nhhvvedagk ihvtlhdgqk ytakivgldp ktdlavikiq

181	aeklpfltfg nsdqlqigdw aiaignpfgl qatvtvgvis akgrnqlhiv dfedfiqtda

241	ainpgnsggp llningqvig vntaivsgsg gyigigfaip slmakrvidq lisdgqvtrg

301	flgvtlqpid selatcykle kvygalvtdv vkgspaekag lrqedvivay ngkeveslsa

361	lrnaislmmp gtrvvlkivr egktieipvt vtqiptedgv salqkmgvrv qnitpeickk

421	lglaadtrgi lvvaveagsp aasagvapgq lilavnrqrv asveelnqvl knskgenvll

481	mvsqgdvvrf ivlksde

CT681 (MOMP, 393 amino acids), ACCESSION# NP 220200.1, GI: 15605414
(SEQ ID NO: 7935)

1	mkkllksvlv faalssassl qalpvgnpae pslmidgilw egfggdpcdp catwcdaism

61	rvgyygdfvf drvlktdvnk efqmgakptt dtgnsaapst ltarenpayg rhmqdaemft

121	naacmalniw drfdvfctlg atsgylkgns asfnlvglfg dnenqktvka esvpnmsfdq

181	svvelytdtt fawsvgaraa lwecgcatlg asfqyaqskp kveelnvlcn aaeftinkpk

241	gyvgkefpld ltagtdaatg tkdasidyhe wqaslalsyr lnmftpyigv kwsrasfdad

301	tiriaqpksa taifdtttln ptiagagdvk tgaegqlgdt mqivslqlnk mksrkscgia

361	vgttivdadk yavtvetrli deraahvnaq frf

CT119 (IncA, 273 amino acids), ACCESSION# NP 219622.1, GI: 15604838
(SEQ ID NO: 7936)

1	mttptlivtp psppapsysa nrvpqpslmd kikkiaaias liligtigfl allghlvgfl

61	iapqitivll alfiislagn alylqktanl hlyqdlqrev gslkeinfml svlqkeflhl

121	skefattskd lsavsqdfys clqgfrdnyk gfeslldeyk nsteemrklf sqeiiadlkg

181	svaslreeir fltplaeevr rlahnqqslt vvieelktir dslrdeigql sqlsktltsq

241	ialqrkessd lcsqiretls sprksaspst kss

CT813 (inclusion membrane protein, 264 amino acids), ACCESSION#
NP 220333.1, GI: 15605547
(SEQ ID NO: 7937)

1	mttlpnncts nsnsintftk diemakqiqg srkdplakts wiaglicvva gvlgllaigi

61	ggcsmasglg ligaivaavi vavglcclvs alclqveksq wwqkefeswi eqksqfrivm

121	admlkanrkl qseveflskg wsddtavhke dvtkyeqvve eyaekimely eetgvltiek

181	inlqkekkaw leekaemeqk lttvtdleaa kqqleekvtd lesekqelre eldkaienld

241	emayeamefe kekhgikpgr rgsi

CT795 (hypothetical protein, 163 amino acids), ACCESSION# NP 220315.1,
GI: 15605529
(SEQ ID NO: 7938)

1	mrfllalfsl ilvlpateaf stedkqcqqe aeedcsqvad tcvfysyaeg lehardegkl

61	tivvlldtsg ysfetladaa hamessllst fadfvvlsrr eavpliyppv pdpmvgeial

121	fleafsdqtf psqpvivtla igassaeimd iteipsinpe fve

CT621 (hypothetical protein, 832 amino acids), ACCESSION# NP 220138.1,
GI: 15605352
(SEQ ID NO: 7939)

1	mnrihrtqgs ltdynstlea iakkiakpds ativsqvaqy eqfkmeqeal kallvsfdqk

61	adqryrnliq rleqldvdrq tgrstesqhi qekpmaslqs enqvvaqavv qsdssmpift

121	gikqswavrl vqgireildq vlvdtslfte eergdllair mdaaslqdkq erlstedirs

181	llslsndvmr vlqkasysst rqleliqsli difgteenle qsfaqvrlen fqailsvike

241	rlteeefrvf qevseeissi qrtseshlsp ehieaiarvg ghlsakives elkasqkvdl

301	cqriaamyqe qvdavqayhs leqdalfvns rqhshfvqvi slvsslmhsl sptseeeril

361	lnpammvsvl ptvraiglrf dfltaeqqqm vnaavsslqq qqldeflgvl cahlvvvncq

421	nketgllegl eesfsetlsg lsnnfvltak mqdilqvcsl qgfvtlangd ryelfsynds

481	geavcdeial gdgfhkvlgt mlavalsqae vfkqecdrfi lqadseknmi hkrmvqgeqk

541	slfltkmqte lnagktiaqt keveasplps avasvlidhy mpkeveflek issrlyygnk

601	gsdigntild aislyvnsat yfgfanyigq ppvvgktren ifagsadnak akldeekkqv

661	dvfleiteaa kttvtnqqsa vtnddklste qkakikaelt qytdmlnais nsltslktql

721	aplsvstveg vdgvfevkng ipgengknwr lvlqtledtv vsgevgsptn igmfqmqalv

781	hlnqqayadm gqnfqlelqm hltsmqqewm vvatslqlln qiylglarnl lr

CT622 (hypothetical protein, 647 amino acids), ACCESSION# NP 220139.1,
GI: 15605353
(SEQ ID NO: 7940)

1	mesgpesyss nqssmnpiin gqiasnsetk estkeseasp sasssvssws flssakhali

61	slrdailnkn ssptdslsql eastststvt rvaardynea ksnfdtaksg lenattlaey

121	etkmadlmaa lqdmerlakq kaevtrikea lqekqevidk lnqlvklekq nqtlketltt

181	tdsadqipai nsqleinkns adqiikdleg qnisyeavlt nagevikass eagiklgqal

241	qsivdagdqs qaavlqaqqn nspdniaatk klidaaetkv nelkqehtgl tdsplvkkae

301	eqisqaqkdi qeikpsgsdi pivgpsgsaa sagsavgalk ssnnsgrisl llddvdnema

361	aiamqgfrsm ieqfnvnnpa takelqamea qltamsdqlv gadgelpaei qaikdalaqa

421	lkqpstdgla tamgqvafaa akvgggsagt agtvqmnvkq lyktafssts sssyaaalsd

481	gysayktlns lysesrsgvq saisqtanpa lsrsysrsgi esqgrsadas qraaetivrd

541	sqtlgdvysr lqvldslmst ivsnpqvnqe eimqkltasi skapqfgypa vqnsadslqk

601	faaqlerefv dgerslaesr enafrkqpaf iqqvlvnias lfsgyls

Claims

What is claimed is:

1. A method of diagnosing tubal factor infertility in a subject, comprising:

(a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof, and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and

(b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and detecting the absence of formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof in the sample, thereby diagnosing tubal factor infertility in the subject.

2. A method of identifying a subject as having an increased likelihood of having or developing tubal factor infertility, comprising:

(a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and

(b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and detecting the absence of formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof in the sample, thereby identifying the subject as having an increased likelihood of having or developing tubal factor infertility.

3. The method of claim 1, wherein the tubal factor infertility is caused by chlamydial infection.

4. The method of claim 1, wherein the subject is a human female.

5. The method of claim 1, wherein the subject is an infertile human female.

6. The method of claim 1, wherein the biological sample is any body fluid in which antibodies can be detected.

7. The method of claim 1, wherein the biological sample is diluted 1:1000.

8. The method of claim 1, wherein the antigen of the CT443 protein or immunologically reactive fragment thereof is a C terminal fragment.

9. The method of claim 1, wherein the diagnostic panel further comprises an additional antigen selected from the group consisting of Chlamydia trachomatis HSP60 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT376 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT557 or an immunologically reactive fragment thereof, or an immunologically reactive fragment thereof and any combination thereof, and an antigen/antibody complex is detected in the sample for each of said additional antigen(s).

10. A method of diagnosing acute Chlamydia trachomatis infection in a subject, comprising:

(b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby diagnosing acute Chlamydia trachomatis infection in the subject.

11. A method of diagnosing acute Chlamydia trachomatis infection in a subject, comprising:

(a) contacting a biological sample from the subject with an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof; and

(b) detecting formation of an antigen/antibody complex with the antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof in the sample, thereby diagnosing acute Chlamydia trachomatis infection in the subject.

12. A method of identifying a subject as having an increased likelihood of having an acute Chlamydia trachomatis infection, comprising:

(b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby identifying a subject as having an increased likelihood of having an acute Chlamydia trachomatis infection.

13. A method of identifying a subject for whom antibiotic therapy for Chlamydia trachomatis infection is likely to be beneficial, comprising: (a) contacting a biological sample from the subject with a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof; and (b) detecting formation of an antigen/antibody complex with each of the antigen of Chlamydia trachomatis CT443 or immunologically reactive fragment thereof, the antigen of Chlamydia trachomatis CT381 or immunologically reactive fragment thereof and the antigen of Chlamydia trachomatis CT875 or the immunologically reactive fragment in the sample, thereby identifying a subject for whom antibiotic therapy to treat Chlamydia trachomatis infection is likely to be beneficial.

14. The method of claim 13, further comprising contacting the sample with an antigen of Chlamydia trachomatis CT147 or immunologically reactive fragment thereof and detecting an antigen/antibody complex in the sample with the antigen of Chlamydia trachomatis CT147 or immunologically reactive fragment thereof.

15. The method of claim 13, wherein the subject is a human female.

16. The method of claim 13, wherein the sample is any body fluid in which antibodies can be detected.

17. A kit comprising a diagnostic panel comprising an antigen of each of Chlamydia trachomatis CT443 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT381 or an immunologically reactive fragment thereof and Chlamydia trachomatis CT875 or an immunologically reactive fragment thereof.

18. The kit of claim 17, wherein the diagnostic panel further comprises an antigen selected from the group consisting of Chlamydia trachomatis HSP60 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT376 or an immunologically reactive fragment thereof, Chlamydia trachomatis CT557 or an immunologically reactive fragment thereof, or an immunologically reactive fragment thereof and any combination thereof.

19. A kit comprising a diagnostic panel comprising an antigen of Chlamydia trachomatis CT875 or immunologically reactive fragment thereof.

20. The kit of claim 17, further comprising an antigen of Chlamydia trachomatis CT147 or immunologically reactive fragment thereof.